Vol. 23 (2018)

DOI: 10.1186/s11658-017-0063-z Volume 23 (2018)
Authors Yanan Li1, Yu Yan2, Fang Liu1, Ming Wang1, Fumin Feng1 and Yonghong Xiao1*
Abstract Background: Our previous study showed that during in vitro experiments changes in calcium concentration were associated with apoptosis. We presumed that the calcium ion might play a role as intermediate messenger for apoptosis-related genes. No such evidence has been reported in the literature. Here, we investigate the effect of calcium ionophore A23187 on the apoptosis of rat hepatic stellate cells (HSCs) stimulated by transforming growth factor-β1 (TGF-β1) to explore the mechanism of apoptosis through the endoplasmic reticulum stress pathway.
Methods: The apoptotic rate was determined using flow cytometry. The changes in Ca2+ level in HSCs were examined with laser confocal microscopy. The expressions of caspase-12 GRP78 and caspase-9 were assayed via western blot.
Results: The respective apoptosis rates for the blank group, the TGF-β1 group and the TGF-β1 + low, medium and high dose calcium ionophore A23187 groups were 3.40 ± 0.10%, 1.76 ± 0.12%, 5.86 ± 0.31%, 11.20 ± 0.48% and 15.08 ± 0.75%, with significant differences between the groups (p < 0.05). The concentration of Ca2+ and the expression of the GRP78, caspase-9 and caspase-12 proteins significantly increased with increasing calcium ionophore A23187 doses (p < 0.05).
Conclusion: Calcium ionophore A23187 increased intracellular Ca2+ and activated endoplasmic reticulum stress, which promoted HSC apoptosis.
Keywords Hepatic fibrosis, Hepatic stellate cells, Calcium ionophore A23187, Apoptosis
Address and Contact Information 1 Department of School of Public Health, North China University of Science and Technology, Hebei, Tang Shan 063000, China.
2 Department of School of Basic Medical Science, North China University of Science and Technology, Hebei,Tang Shan 063000, China.
* Corresponding author: 1619747081@qq.com
Read full article at BMC

DOI: 10.1186/s11658-017-0058-9 Volume 23 (2018)
Authors Ying Chen1,2,3*, Lei Zhang1,2,3, Wen-xin Liu1 and Ke Wang1
Abstract Background: Anti-angiogenesis therapy that targets VEGF is one of the important treatment strategies in advanced ovarian cancer. However, depending on the pharmaceutical agent, treatment can have undesirable side effects. SEMA4D has recently gained interest for its role in promoting angiogenesis. Here, we try to further understand the mechanism by which SEMA4D promotes angiogenesis in ovarian cancer.
Methods: Correlation and western blot assaya were used to detect the relationship between VEGF and SEMA4D in clinical tissues and cells. Vasculogenic mimicry and transwell migration analyses were used to detect the roles of VEGF, SEMA4D and plexin-B1 on vasculogenic mimicry and migration. Vascular density and SEMA4D expression was determined using immunofluorescence staining in clinical tissues of EOC. Western blot was used to detect the expressions of CD31, MMP2 and VE-cadherin. We also analyzed the relationship between VEGF-SEMA4D and malignant tumor prognosis.
Results: We found that knockdown of VEGF could suppress SEMA4D expression and that the expressions of VEGF and SEMA4D have a positive correlation in EOC cancer tissues. Vasculogenic mimicry and transwell migration analyses showed that SEMA4D and VEGF have a synergistic effect on the promotion of angiogenesis in A2780 and HUVEC cells. Soluble SEMA4D (sSEMA4D) could promote VM and migration in A2780 and HUVEC cells via the SEMA4D/plexin-B1 pathway, but the effect was not noted in stably transfected shR-plexin-B1 cells. In clinical tissues of EOC, the vascular density and SEMA4D/plexin-B1 expression were higher. When VEGF, SEMA4D and plexin-B1 was knocked down, the expression of CD31, MMP2 and VE-cadherin, which are the markers and initiators of angiogenesis and the epithelial–mesenchymal transition (EMT) process were reduced. VEGF and SEMA4D had a positive correlation with the malignant degree of ovarian cancer, and SEMA4D can serve as an independent prognostic factor.
Conclusions: VEGF and SEMA4D have synergistic effects on the promotion of angiogenesis in epithelial ovarian cancer. Targeting VEGF and the SEMA4D signaling pathway could be important for the therapy for EOC.
Keywords SEMA4D, Vegf, Angiogenesis, Tumor growth, Epithelial ovarian cancer
Address and Contact Information 1Department of Gynecologic Oncology, Tianjin Medical University Cancer Institute and Hospital, Huanhuxi Road, Hexi District, Tianjin 300060, China.
2 Key Laboratory of Cancer Prevention and Therapy, Tianjin 300060, China.
3 National Clinical Research Centre of Cancer, Tianjin 300060, China.
* Corresponding author: chenying912a@126.com
Read full article at BMC

DOI: 10.1186/s11658-017-0068-7 Volume 23 (2018)
Authors Paul Chen, Saifuddin Sheikh, Ateeq Ahmad, Shoukath M. Ali, Moghis U. Ahmad and Imran Ahmad*
Abstract Endoxifen, an active metabolite of tamoxifen, has been shown to be an effective anti-estrogenic agent in estrogen receptor-positive breast cancer patients. In melanoma, estrogen receptor expression is shown to be associated with disease progression. However, the therapeutic benefit of endoxifen in melanoma has not yet been evaluated. Here, we present the first demonstration of the anti-melanogenic activity of endoxifen in vitro and in vivo.
The in vitro cytotoxic effect of endoxifen was tested using a cell viability assay. The in vivo anti-melanogenic activity was evaluated in B16F10 cell-bearing C57BL/6 mice, a mouse melanoma model. The general toxicity was tested in Swiss albino mice. Endoxifen exhibited greater activity against melanoma cell lines. Treatment of B16F10 mouse and SK-MEL-5 human melanoma cell lines with 10 μM of endoxifen for 48 h respectively resulted in 93.6 and 92.5% cell death. Orally administered endoxifen, at dose levels of 4 and 8 mg/kg body weight/day for 20 consecutive days, respectively reduced metastatic melanoma nodules in the lungs by 26.7 and 82.7%.
Endoxifen was found to be a safe and effective anti-melanogenic agent in animal studies.
Keywords Endoxifen, Melanoma tumor model, Tamoxifen, Safety, Efficacy
Address and Contact Information Jina Pharmaceuticals, Inc., 28100 N. Ashley Circle, Suite 103, Libertyville, IL 60048, USA
* Corresponding author: imran@jinapharma.com
Read full article at BMC

DOI: 10.1186/s11658-018-0071-7 Volume 23 (2018)
Authors Shaohua Wang1*, Hongxing Dang2, Feng Xu2, Jian Deng1 and Xuemei Zheng1
Abstract Background: Calcitonin gene-related peptide (CGRP) can protect against hyperoxia-induced lung injury, making the upregulation of CGRP a potential therapeutic approach for this type of injury. However, the effects of CGRP on the Wnt7b/β-catenin signaling pathway are unclear. In this study, we investigated the roles of CGRP and the Wnt7b/β-catenin signaling pathway in hyperoxia-induced lung injury.
Methods: Premature Sprague Dawley (SD) rats were exposed to 21, 40, 60 and 95% oxygen for 3, 7 and 14 days. The animals’ body weights, survival rates and endogenous CGRP levels were measured. Lung samples were harvested for histological analyses and measurements of malondialdehyde (MDA) concentration and total antioxidant capacity (TAOC). We also assessed the MDA concentration and TAOC in the lung tissues after administration of 200 nmol/kg CGRP8–37 (a CGRP antagonist). Finally, alveolar epithelial type II (AEC II) cells were isolated from premature rats, exposed to 21 or 95% oxygen for 3, 7 and 14 days, and treated with 10−8 mol/l exogenous CGRP. The protein expressions of Wnt7b and β-catenin were assessed using western blotting, and TCF and c-myc mRNA expressions were assessed using qPCR.
Results: Rats exposed to 60 and 95% oxygen had significantly lower body weights and survival rates than the 21 and 40% groups, and the decrease was time dependent. Endogenous CGRP was elevated in the lung tissues of premature rats exposed to 95% oxygen. CGRP8–37 induced apparent inflammation in the lung tissue and alveolar structural remodeling. In addition, the expression levels of Wnt7b and β-catenin were markedly increased after exposure for 3 days. They peaked at 7 days, then declined at 14 days. The levels of TCF/c-myc in AEC II cells increased significantly after CGRP treatment when compared with cells that had only undergone hyperoxia.
Conclusions: CGRP protected against hyperoxia-induced lung injury in premature rats. This process involves the Wnt7b/β-catenin signaling pathway.
Keywords CGRP, Lung injury, Wnt7b, β-catenin
Address and Contact Information 1 Neonatal Intensive Care Unit, Women and Children Health Institute of Futian, University of South China, Jintian South Road No. 2002, Futian district, Shen Zhen 518045, China.
2 Pediatric Intensive Care Unit, Children’s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, Yu Zhong, Chongqing 400014, China.
* Corresponding author: wangshua2017@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0073-5 Volume 23 (2018)
Authors Nazli F. Eskici1, Sevim Erdem-Ozdamar2 and Didem Dayangac-Erden1*
Abstract Background: Perineuronal nets (PNNs), which are localized around neurons during development, are specialized forms of neural extracellular matrix with neuroprotective and plasticity-regulating roles. Hyaluronan and proteoglycan link protein 1 (HAPLN1), tenascin-R (TNR) and aggrecan (ACAN) are key elements of PNNs. In diseases characterized by neuritogenesis defects, the expression of these proteins is known to be downregulated, suggesting that PNNs may have a role in neural differentiation.
Methods: In this study, the mRNA and protein levels of HAPLN1, TNR and ACAN were determined and compared at specific time points of neural differentiation. We used PC12 cells as the in vitro model because they reflect this developmental process.
Results: On day 7, the HAPLN1 mRNA level showed a 2.9-fold increase compared to the non-differentiated state. However, the cellular HAPLN1 protein level showed a decrease, indicating that the protein may have roles in neural differentiation, and may be secreted during the early period of differentiation. By contrast, TNR mRNA and protein levels remained unchanged, and the amount of cellular ACAN protein showed a 3.7-fold increase at day 7. These results suggest that ACAN may be secreted after day 7, possibly due to its large amount of post-translational modifications.
Conclusions: Our results provide preliminary data on the expression of PNN elements during neural differentiation. Further investigations will be performed on the role of these elements in neurological disease models.
Keywords Perineuronal nets, HAPLN1, Tenascin-R, Aggrecan, PC12 differentiation
Address and Contact Information 1 Faculty of Medicine Department of Medical Biology, Hacettepe University, Ankara, Turkey. 2Faculty of Medicine Department of Neurology, Hacettepe University, Ankara, Turkey.
* Corresponding author: didayan@hacettepe.edu.tr
Read full article at BMC

DOI: 10.1186/s11658-017-0067-8 Volume 23 (2018)
Authors Guodong Li1†, Xiaoling Hu2†, Lu Sun1, Xin Li1, Jianfeng Li1, Tongli Li1* and Xiaohui Zhang3*
Abstract Background: Laryngeal cancer tends to have a very poor prognosis due to the unsatisfactory efficacy of chemotherapy for this cancer. Multidrug resistance (MDR) is the main cause of chemotherapy failure. The proto-oncogene c-fos has been shown to be involved in the development of MDR in several tumor types, but few studies have evaluated the relationship between c-fos and MDR in laryngeal cancer. We investigated the role of c-fos in MDR development in laryngeal cancer cells (cell line: human epithelial type 2, HEp-2) using the chemotherapeutic vincristine (VCR).
Methods: HEp-2/VCR drug resistance was established by selection against an increasing drug concentration gradient. The expressions of c-fos and multidrug resistance 1 (mdr1) were measured using qPCR and western blot. C-fos overexpression or knockdown was performed in various cells. The intracellular rhodamine-123 (Rh-123) accumulation assay was used to detect the transport capacity of P-glycoprotein (P-gp, which is encoded by the mdr1 gene).
Results: HEp-2 cells with VCR-induced resistance (HEp-2/VCR cells) were not only resistant to VCR but also evolved cross-resistance to other chemotherapeutic drugs. The expressions of the c-fos and mdr1genes were significantly higher in the HEp-2/VCR cells than in control cells. C-fos overexpression in HEp-2 cells (c-fos WT) resulted in increased P-gp expression and increased the IC50 for 5-FU. C-fos knockdown in the HEp-2/VCR cells (c-fos shRNA) resulted in decreased P-gp expression and decreased IC50 for 5-FU. An intracellular Rh-123 accumulation assay showed that the mean intracellular fluorescence intensity (MFI) was lower in the HEp-2/VCR cells than in HEp-2 cells. C-fos WT cells also showed lower MFI. By contrast, c-fos shRNA cells exhibited a higher MFI than the control group.
Conclusion: C-fos increased the expression of P-gp and mdr1 in the HEp-2/VCR cells, and enhanced the efflux function of the cells, thereby contributing to the development of MDR.
Keywords C-fos, P-glycoprotein, Multidrug resistance, Laryngeal carcinoma
Address and Contact Information 1 Department of Otorhinolaryngology, Shanxi Provincial People’s Hospital Affiliated to Shanxi Medical University, Taiyuan, Shanxi 030012, China.
2 Department of Pharmacology, Shanxi Medical University, Taiyuan, Shanxi, China.
3 Artificial Livers Treatment Center, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China.
* Corresponding author: t.l.li@163.com;zhangxiaohui_111@163.com
Equal contributors
Read full article at BMC

DOI: 10.1186/s11658-018-0072-6 Volume 23 (2018)
Authors Hao Yang1, Di Wu1, Hua Li1, Nan Chen1 and Yongjun Shang2*
Abstract Background: Osteoarthritis is characterized by the continuous degradation of the articular cartilage. The microRNA miR-448 has been found to be broadly involved in cellular processes, including proliferation, apoptosis, invasion and EMT. While aberrant expression of miR-448 has been found in multiple cancers, its level in osteoarthritis cartilage and its role in the progression of this disease are still unknown. Here, we examined the functional roles of miR-448 and its expression in osteoarthritis tissues, including IL-1β-stimulated osteoarthritis chondrocytes.
Methods: Chondrocytes were isolated from human articular cartilage and stimulated with IL-1β. The expression levels of miR-448 in the cartilage and chondrocytes were both determined. After transfection with an miR-448 mimic or inhibitor, the mRNA levels of aggrecan, type II collagen and MMP-13 were determined. Luciferase reporter assay, qRT-PCR and western blot were performed to explore whether matrilin-3 was a target of miR-448. Furthermore, we co-transfected chondrocytes with miR-448 inhibitor and siRNA for matrilin-3 and then stimulated them with IL-1β to determine whether miR-448-mediated IL-1β-induced cartilage matrix degradation resulted from directly targeting matrilin-3.
Results: The level of miR-448 was significantly higher and matrilin-3 expression was significantly lower in osteoarthritis cartilage and IL-1β-induced chondrocytes than in normal tissues and cells. Furthermore, matrilin-3 expression was reduced by miR-448 overexpression. MiR-448 downregulation significantly alleviated the IL-1β-induced downregulation of aggrecan and type II collagen expression, and upregulation of MMP-13 expression. MiR-448 overexpression had the opposite effects. Knockdown of matrilin-3 reversed the effects of the miR-448 inhibitor on the expressions of aggrecan, type II collagen and MMP-13.
Conclusion: The findings showed that miR-448 contributed to the progression of osteoarthritis by directly targeting matrilin-3. This indicates that it has potential as a therapeutic target for the treatment of osteoarthritis.
Keywords Osteoarthritis, MicroRNA-448, Interleukin-1 beta, Chondrocyte, Extracellular matrix, Matrilin-3
Address and Contact Information 1 Department of Orthopedics, First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan Province, People’s Republic of China.
2 Department of Orthopedics, Dalian University Affiliated Xinhua Hospital, No. 156 Xinhua Street, Shahekou District, Dalian 116021, Liaoning Province, People’s Republic of China.
* Corresponding author: shangyongjundalian@yahoo.com
Read full article at BMC

DOI: 10.1186/s11658-018-0075-3 Volume 23 (2018)
Authors Tomoaki Kinjo1,3, Chuanhai Sun1,4, Tomomi Ikeda2, Takako Ikegami2, Yuhki Tada1,5, Tadayuki Akagi1, Takashi Yokota1 and Hiroshi Koide1,2*
Abstract Background: Platelet-derived growth factor-C (PDGF-C) has been shown to be involved in several biological processes, such as embryonic development, wound healing and angiogenesis, as well as in diseases including tumor formation and fibrotic diseases. However, its role in fibrosarcoma and embryonic stem (ES) cells has not been elucidated.
Methods: The expression level of PDGF-C was measured using RT-PCR. The activity of PDGF-C was suppressed using RNA interference or a neutralizing antibody and the effect on cell growth was examined using the WST and soft agar assays.
Results: In the tumor cell lines studied, the highest level of PDGF-C expression was in human HT1080 fibrosarcoma cells. In ES cells, it was highly expressed in the self-renewal state but not in the differentiated state. PDGF-C knockdown suppressed anchorage-dependent and -independent growth of HT1080 and ES cells. In addition, the suppression of PDGF-C activity by a neutralizing antibody retarded ES cell growth.
Conclusion: Our results suggest that PDGF-C plays an important role in the proliferation of fibrosarcoma and ES cells.
Keywords Embryonic stem cells, Cancer, PDGF-C, Fibrosarcoma, Anchorage-independent growth
Address and Contact Information 1Department of Stem Cell Biology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Ishikawa, Japan.
2 Laboratory of Molecular and Biochemical Research, Research Support Center, Juntendo University Graduate School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
3 Present address: Department of Diagnostic Imaging and Nuclear Medicine, Graduate School of Medicine, Kyoto University, Sakyo-ku, Kyoto, Japan.
4 Present address: Neusoft Xikang Healthcare Technology Co., Ltd., Shenyang, China.
5 Present address: RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
* Corresponding author: h-koide@juntendo.ac.jp
Read full article at BMC

DOI: 10.1186/s11658-018-0076-2 Volume 23 (2018)
Authors Seungwoo Kim1, Hyosoon Cheon1, Sam-Moon Kim1 and Young-Youl Kim2*
Abstract Background: Previous studies indicated that cadmium (Cd) increases PI3-kinase/Akt phosphorylation, resulting in an alteration in GSK-3β activity. However, the mechanism of Cd-induced endoplasmic reticulum (ER) stress in neuronal cells has yet to be studied in needs further elucidation. We examined the role of GSK-3β in Cd-induced neuronal cell death and the related downstream signaling pathways.
Methods: SH-SY5Y human neuroblastoma cells were treated with 10 or 20 μM BAPTA-AM and 1 μM wortmannin for 30 min and then incubated with 25 μM Cd for 12 h. Apoptotic cells were visualized via DAPI and PI staining. Data were evaluated with one-way analysis of variance (ANOVA) followed by Student’s t-test. Data are expressed as the means ± SD of experiments performed at least three times.
Results: Treatment of human neuronal SH-SY5Y cells with Cd induced ER, stress as evidenced by the increased expression of GRP78, which is a marker of ER stress. Cd exposure significantly increased the phosphorylation of Akt at thr308 and ser473 and that of GSK-3β at ser9 in a time-dependent manner, while the total protein levels of GSK-3β and Akt did not change. Cd-induced apoptosis was higher in GSK-3β-knockdown cells than in normal cells.
Conclusions: Our data suggest that Akt/GSK-3β signaling activated by Cd is involved in neuronal cell survival.
Keywords Cadmium, ER-stress, GSK-3β
Address and Contact Information 1 Division of Brain Diseases, Center for Biomedical Science, National Institute of Health, Center for Disease Control & Prevention, Osong Health Technology Administration Complex, 187, Osongsaengmyeong2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, South Korea.
2 Division of Biobank for Health Sciences, Center for Genome Science, National Institute of Health, Center for Disease Control & Prevention, 200 Osongsaengmyeong2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, South Korea.
* Corresponding author: youngyk@nih.go.kr
Read full article at BMC

DOI: 10.1186/s11658-018-0080-6 Volume 23 (2018)
Authors Ghmkin Hassan1*, Mohammad Bahjat2, Issam Kasem2,3, Chadi Soukkarieh2,3 and Majd Aljamali1,2,3*
Abstract Purpose: Articular cartilage has a poor capacity for self-repair, and thus still presents a major challenge in orthopedics. Mesenchymal stem cells (MSCs) are multipotent stem cells with the potential to differentiate into chondrocytes in the presence of transforming growth factor beta (TGF-β). Platelet lysate (PL) contains a relatively large number of growth factors, including TGF-β, and has been shown to ameliorate cartilage repair. Here, we investigated the ability of PL to direct chondrogenic differentiation of MSCs along with other standard differentiation components in a pellet culture system.
Methods: We isolated and expanded MSCs from human umbilical cords using a PL-supplemented medium and characterized the cells based on immunophenotype and potential for differentiation to adipocytes and osteocytes. We further cultured MSCs as pellets in a chondrogenic-differentiation medium supplemented with PL. After 21 days, the pellets were processed for histological analysis and stained with alician blue and acridine orange. The expression of SOX9 was investigated using RT-PCR.
Results: MSCs maintained their stemness characteristics in the PL-supplemented medium. However, the distribution of cells in the pellets cultured in the PL-supplemented chondrogenic differentiation medium had a greater similarity to cartilage tissue-derived chondrocytes than to the negative control. The intense alician blue staining indicated an increased production of mucopolysaccharides in the differentiated pellets, which also showed elevated expression of SOX9.
Conclusions: Our data suggest that MSCs could be differentiated to chondrocytes in the presence of PL and absence of exogenous TGF-β. Further research needs to be conducted to understand the exact role and potential of PL in chondrogenic differentiation and chondrocyte regeneration.
Keywords Platelet lysate, Mesenchymal stem cells, Chondrogenic differentiation, Cartilage
Address and Contact Information 1 Faculty of Pharmacy, Damascus University, Damascus, Syria.
2 Faculty of Sciences, Damascus University, Damascus, Syria.
3 National Commission for Biotechnology (NCBT), Damascus, Syria.
* Corresponding author: hsn.ghmkin@gmail.com; maljamali@gmail.com
Read full article at BMC

DOI: 10.1186/s11658-018-0079-z Volume 23 (2018)
Authors Laura Susan Boyd1, Devrim Gozuacik2 and Anna Margaretha Joubert1*
Abstract Background: The majority of novel chemotherapeutics target the cell cycle, aiming to effect arrest and cause apoptosis. One such agent, 2-methoxyestradiol (2ME), has been shown to possess anticancer properties against numerous cancer types, both in vitro and in vivo. Despite its promise, 2ME has exhibited limitations, including low oral bioavailability and rapid hepatic enzymatic inactivation in vivo. A novel sulphamoylated estrogen analog, 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16), was in silico-designed in our laboratory to overcome these issues. It was then synthesized by a pharmaceutical company and used in an in vitro antiproliferative effect study on a human cervical carcinoma (HeLa) cell line.
Results: Cell proliferation data obtained from the crystal violet assay and real-time cell analysis demonstrated that 0.2 μM of ESE-16 had a significant inhibitory effect on the HeLa cells 24 h post-exposure. Immunofluorescence showed that ESE-16 is a microtubule disruptor that causes cells to undergo a mitotic block. Qualitative morphological studies using polarization-optical transmitted light differential interference contrast (PlasDIC) and light microscopy revealed a decrease in cell density and an increase in the number of cells arrested in metaphase. After ESE-16 exposure, hallmarks of apoptosis were also observed, including membrane blebbing, chromatin condensation and the presence of apoptotic bodies. Flow cytometry provided quantitative results from cell cycle progression analysis, indicating cells undergoing apoptosis and cells in the G2/M phase of the cell cycle, confirming cell cycle arrest in metaphase after ESE-16 treatment. Quantification of the ESE-16-mediated upregulation of cyclin B in HeLa cells and spectrophotometric and flow cytometric confirmation of cell death via apoptosis further confirmed the substance’s impact.
Conclusion: ESE-16 exerts its antiproliferative effects through microtubule disruption, which induces a mitotic block culminating in apoptosis. This research provided information on ESE-16 as a potential antitumor agent and on cellular targets that could aid in the design of prospective microtubule-disrupting compounds. Further in vitro and in vivo investigations of this novel compound are needed.
Keywords Estradiol analog, ESE-16, Antiproliferative, Metaphase block, Cervical carcinoma
Address and Contact Information 1Department of Physiology, Faculty of Health Sciences, University of Pretoria, Private Bag X323, Arcadia 0007, Pretoria, South Africa.
2 Faculty of Engineering and Natural Sciences, Molecular Biology Genetics and Bioengineering Program, Sabanci University, Orhanli-Tuzla 3495, Istanbul, Turkey.
* Corresponding author: annie.joubert@up.ac.za
Read full article at BMC

DOI: 10.1186/s11658-018-0078-0 Volume 23 (2018)
Authors Michaela Galoczova*, Philip Coates and Borivoj Vojtesek
Abstract Signal Transducer and Activator of Transcription 3 (STAT3) is a transcription factor with many important functions in the biology of normal and transformed cells. Its regulation is highly complex as it is involved in signaling pathways in many different cell types and under a wide variety of conditions. Besides other functions, STAT3 is an important regulator of normal stem cells and cancer stem cells. p63 which is a member of the p53 protein family is also involved in these functions and is both physically and functionally connected with STAT3. This review summarizes STAT3 function and regulation, its role in stem cell and cancer stem cell properties and highlights recent reports about its relationship to p63.
Keywords STAT3, Stem cells, Cancer stem cells, p63
Address and Contact Information Regional Centre for Applied Molecular Oncology, Masaryk Memorial Cancer Institute, Zluty kopec 7, 656 53 Brno, Czech Republic
* Corresponding author: michaela.galoczova@mou.cz
Read full article at BMC

DOI: 10.1186/s11658-018-0074-4 Volume 23 (2018)
Authors Yuanyuan Sun1, Chen Wang2, Ligang Wang2, Zhibo Dai2 and Kongbin Yang2*
Abstract Background: Arsenic trioxide (As2O3) has a dramatic therapeutic effect on acute promyelocytic leukemia (APL) patients. It can also cause apoptosis in various tumor cells. This study investigated whether As2O3 has an antitumor effect on glioma and explored the underlying mechanism.
Results: MTT and trypan blue assays showed that As2O3 remarkably inhibited growth of C6 and 9 L glioma cells. Cell viability decreased in glioma cells to a greater extent than in normal glia cells. The annexin V-FITC/PI and Hoechest/PI staining assays revealed a significant increase in apoptosis that correlated with the duration of As2O3 treatment and occurred in glioma cells to a greater extent than in normal glial cells. As2O3 treatment induced reactive oxygen species (ROS) production in C6 and 9 L cells in a time-dependent manner. Cells pretreated with the antioxidant N-acetylcysteine (NAC) showed significantly lower As2O3-induced ROS generation. As2O3 significantly inhibited the expression of the anti-apoptotic gene Bcl-2, and upregulated the proapoptotic gene Bax in both C6 and 9 L glioma cells in a time-dependent manner.
Conclusions: As2O3 can significantly inhibit the growth of glioma cells and it can induce cell apoptosis in a time- and concentration-dependent manner. ROS were found to be responsible for apoptosis in glioma cells induced by As2O3. These results suggest As2O3 is a promising agent for the treatment of glioma.
Keywords Arsenic trioxide (As2O3), Reactive oxygen species (ROS), Glioma, Apoptosis
Address and Contact Information 1 Nursing Support Center, First Affiliated Hospital, Harbin Medical University, Harbin 150000, China.
2 Neurosurgery Department, First Affiliated Hospital, Harbin Medical University, Nangang District, Harbin 150000, China.
* Corresponding author: dk5732@163.com; ykbnewrosurgery@sina.com
Read full article at BMC

DOI: 10.1186/s11658-018-0081-5 Volume 23 (2018)
Authors Qingyun Lei, Jian Tan, Shangqing Yi, Na Wu, Yilin Wang and Heng Wu*
Abstract Background: The regulation of microglial function via mitochondrial homeostasis is important in the development of neuroinflammation. The underlying mechanism for this regulatory function remains unclear. In this study, we investigated the protective role of mitochonic acid 5 (MA-5) in microglial mitochondrial apoptosis following TNFα-induced inflammatory injury.
Methods: TNFα was used to induce inflammatory injury in mouse microglial BV-2 cells with and without prior MA-5 treatment. Cellular apoptosis was assessed using the MTT and TUNEL assays. Mitochondrial functions were evaluated via mitochondrial membrane potential JC-1 staining, ROS flow cytometry analysis, mPTP opening assessment, and immunofluorescence of cyt-c. Mitophagy was examined using western blots and immunofluorescence. The pathways analysis was carried out using western blots and immunofluorescence with a pathway blocker.
Results: Our results demonstrated that TNFα induced apoptosis in the microglial BV-2 cell line by activating the caspase-9-dependent mitochondrial apoptotic pathway. Mechanistically, inflammation reduced mitochondrial potential, induced ROS production, and contributed to the leakage of mitochondrial pro-apoptotic factors into the cytoplasm. The inflammatory response reduced cellular energy metabolism and increased oxidative stress. By contrast, treatment with MA-5 reduced mitochondrial apoptosis via upregulation of mitophagy. Increased mitophagy degraded damaged mitochondria, disrupting mitochondrial apoptosis, neutralizing ROS overproduction, and improving cellular energy production. We also identified that MA-5 regulated mitophagy via Bnip3 through the MAPK–ERK–Yap signaling pathway. Inhibiting this signaling pathway or knocking down Bnip3 expression prevented MA-5 from having beneficial effects on mitochondrial homeostasis and increased microglial apoptosis.
Conclusions: After TNFα-induced inflammatory injury, MA-5 affects microglial mitochondrial homeostasis in a manner mediated via the amplification of protective, Bnip3-related mitophagy, which is mediated via the MAPK–ERK–Yap signaling pathway.
Keywords MA-5, Inflammatory injury, Mitophagy, Microglia, Mitochondria, MAPK–ERK– yap signaling pathway
Address and Contact Information Department of Neurology, First Hospital Affiliated to University of South China, Hunan, China
* Corresponding author: jiantan198@hotmail.com
Equal contributors
Read full article at BMC

DOI: 10.1186/s11658-018-0083-3 Volume 23 (2018)
Authors Ruijie Gong1, Xiaofei Lv3 and Fengqiong Liu1,2*
Abstract Background: The miRNA cluster miR-17-92 is known to act as an oncogene in various cancers. Members of this cluster were also found to be involved in some other pathological process, such as steatosis, which is a pivotal event in the initiation and progression of nonalcoholic fatty liver disease (NAFLD). This study aimed to explore whether miR-17, one of the most functional miRNAs in the miR-17-92 family, participates in the process of steatosis in hepatoma cells.
Methods: We developed both a miR-17-expressing transgenic mouse model and a miR-17-expressing HepG2 cell model, the latter was established via stable transfection. Real-time PCR and western blot were applied to measure the expression levels of miR-17 and the potential target gene CYP7A1. The luciferase assay was used to confirm direct binding of miR-17 and CYP7A1. The oleic acid induction assay and Oil-Red-O staining were performed to support the determination of steatotic changes in HepG2 cell.
Results: Extensive steatotic changes were observed in the livers of transgenic mice. Fewer were seen in the wild-type animals. CYP7A1 was confirmed as a target gene of miR-17, and the expression of CYP7A1 was found to be negatively regulated in both the transgenic mice liver cells and the miR-17-expressing HepG2 cells. CYP7A1 was found to participate in miR-17-induced steatosis, as its repressed expression in miR-17 HepG2 cells exacerbated steatotic change. Re-introduction of CYP7A1 into miR-17 HepG2 cell partially alleviated steatosis.
Conclusions: miR-17 is a novel regulator of CYP7A1 signaling in hepatic lipid metabolism, suggesting a potential therapeutic approach for fatty liver.
Keywords MiRNA-17, Steatosis, Fatty liver, CYP7A1
Address and Contact Information 1Fujian Provincial Key Laboratory of Environment Factors and Cancer, School of Public Health, Fujian Medical University, Fuzhou, Fujian 350008, People’s Republic of China.
2 Department of Epidemiology and Health Statistics, School of Public Health, Fujian Medical University, Fujian, China.
3 Department of Internal Medicine, Guangdong Women and Children’s Hospital, Guangzhou Medical University, 521 Xingnan Road, Guangzhou, China.
* Corresponding author: meimei20061986@126.com
Read full article at BMC

DOI: 10.1186/s11658-018-0082-4 Volume 23 (2018)
Authors Gabriela Sánchez1, Omar Estrada1, Giovana Acha1, Alfonso Cardozo2, Franshelle Peña1, Marie Christine Ruiz1, Fabián Michelangeli1 and Claudia Alvarado-Castillo1,3*
Abstract Background: The leaves of Annona purpurea have yielded several alkaloids with anti-aggregation activities against rabbit platelets. This is promising in the search for agents that might act against platelets and reduce the incidence of cardiovascular diseases. Since significant differences in platelet function have been reported between human and animal platelets, a study focusing on the effect of A. purpurea extracts against human platelet activation is necessary.
Methods: The compounds in an A. purpurea ethanolic extract underwent bio-guided fractionation and were used for in vitro human platelet aggregation assays to isolate the compounds with anti-platelet activity. The bioactive compounds were identified by spectroscopic analysis. Additional platelet studies were performed to characterize their action as inhibitors of human platelet activation.
Results: The benzylisoquinoline alkaloid norpurpureine was identified as the major anti-platelet compound. The IC50 for norpurpureine was 80 μM against platelets when stimulated with adenosine 5′-diphosphate (ADP), collagen and thrombin. It was pharmacologically effective from 20 to 220 μM. Norpurpureine (220 μM) exhibited its in vitro effectiveness in samples from 30 healthy human donors who did not take any drugs during the 2 weeks prior to the collection. Norpurpureine also gradually inhibited granule secretion and adhesion of activated platelets to immobilized fibrinogen. At the intra-platelet level, norpurpureine prevented agonist-stimulated calcium mobilization and cAMP reduction. Structure–activity relationship analysis indicates that the lack of a methyl group at the nitrogen seems to be key in the ability of the compound to interact with its molecular target.
Conclusion: Norpurpureine displays a promising in vitro pharmacological profile as an inhibitor of human platelet activation. Its molecular target could be a common effector between Ca2+ and cAMP signaling, such as the PLC-PKC-Ca2+ pathway and PDEs. This needs further evaluation at the protein isoform level.
Keywords Annona purpurea, Alkaloids, Norpurpureine, Human platelets
Address and Contact Information 1 Centro de Biofísica y Bioquímica (CBB), Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Bolivarian Republic of Venezuela.
2 Laboratorio de Botánica Sistemática, Facultad de Agronomía, Universidad Central de Venezuela (UCV), Maracay, Bolivarian Republic of Venezuela.
3 Laboratorio de Hemostasia y Genética Vascular, Centro de Biofísica y Bioquímica, Instituto Venezolano de Investigaciones Científicas, Apartado 20632, K11 de la Carretera Panamericana, Caracas 1020-A, Bolivarian Republic of Venezuela.
* Corresponding author: cpilar.alvarado@gmail.com; calvarad@ivic.gob.ve
Read full article at BMC

DOI: 10.1186/s11658-018-0070-8 Volume 23 (2018)
Authors Jing Liu1, Chunxia Yang1, Yufang Gu2, Chong Li1, Huamei Zhang1, Wenfang Zhang1, Xueqing Wang1, Nan Wu1 and Chunyan Zheng3*
Abstract Background:, Epstein–Barr virus (EBV) infection is causatively associated with a variety of human cancers, including gastric cancer (GC), which has one of the highest mortality rates of all human cancers. Long non-coding RNAs (lncRNAs) show important regulatory roles in human GC. SNHG8 is a recently identified lncRNA that was reported to show abnormal expression pattern in GC. However, little is known of its biological function in EBV-associated GC.
Methods: We used cell viability, colony formation and cell cycle assays to investigate the roles of lncRNA SNHG8 in the cell growth of EBV-associated GC.
Results: The transcript levels of SNHG8 in the cultured EBV-associated GC cells were significantly higher in the cultured EBV-associated GC cells compared with the levels in normal human gastric mucosal cells and EBV-negative GC cells. Knockdown of SNHG8 with specific shRNAs inhibited cell proliferation and colony formation and arrested the cell cycle in the G0/G1 phase in vitro. We also found that knockdown of SNHG8 suppressed tumor growth in vivo.
Conclusions: These data indicate the pro-oncogenic potential of SNHG8 in EBV-associated GC, meaning it is a latent therapeutic target for the treatment of this type of cancer.
Keywords SNHG8, Cell growth, shRNA, Epstein-Barr virus-associated gastric carcinoma
Address and Contact Information 1 Department of Nephology, Zibo, China.
2 Department of Gastrointestinal Surgery, Zibo, China.
3 Department of Medical Care, Zibo Central Hospital, 54 Gongqingtuan Rd, Zibo, Shandong 255036, People’s Republic of China.
* Corresponding author: Chunyanzheng2017@163.com
Read full article at BMC

DOI: /10.1186/s11658-018-0084-2 Volume 23 (2018)
Authors Bihua Liang1†, Liqian Peng1,2†, Runxiang Li1, Huaping Li1, Ziyin Mo1,2, Xinyue Dai1, Na Jiang3, Qing Liu4, Erting Zhang5, Huiyan Deng1, Zhenjie Li1* and Huilan Zhu1*
Abstract Background: Lycium barbarum polysaccharide (LBP) is considered an antioxidant agent. NF-E2-related factor-2 (Nrf2) is an important regulator for protection against UV damage. In this study, we verified the performance of LBP and the correlation between LBP and Nrf2.
Methods: HSF cells were treated with LBP to determine dose and time dependencies. An antioxidant response element (ARE) reporter was designed to monitor the activity of the Nrf2 antioxidant pathway.
Results: For HSF cells, the optimal LBP treatment was 300 μg/ml for 3 h. The ARE-reporter assay showed that LBP could increase the robustness of p-Nrf2. Treatments with genistein and LY294002 reduced of nuclear p-Nrf2 after 24 h. LBP increased the level of nuclear Nrf2, which functions by both phosphorylation and nuclear translocation. Silencing Nrf2 led to increased reactive oxygen species (ROS) levels, lower cell viability, and decreased superoxide dismutase (SOD) and glutathione peroxidase (GSP-PX) levels. This induced a higher level of lipid peroxide (LPO). However, LBP could decrease the levels of ROS and LPO and enhance the levels of SOD and GSP-PX.
Conclusion: LBP protects HSF cells against UV damage via the regulation of Nrf2.
Keywords Ultraviolet damage, Lycium barbarum, Polysaccharide, Nrf2
Address and Contact Information 1 Guangzhou Institute of Dermatology, Guangzhou, China.
2 Guangzhou Medical University, Guangzhou, China.
3 Guangzhou First People’s Hospital, Guangzhou, China.
4 The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
5 The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
* Corresponding author: lizhen3564@163.com; zhlhuilan@126.com
Equal contributors
Read full article at BMC

DOI: 10.1186/s11658-018-0086-0 Volume 23 (2018)
Authors Renguo Xu1, Zhen Liu1, Jiande Hou1, Tao Huang2 and Ming Yang1*
Abstract Background: Osthole is a natural product that has multiple bioactive functions and has been reported to exert potent immunosuppressive effects. However, the therapeutic effect of osthole on arthritis has not been explored. In the present study, a collagen-induced arthritis rat model, IL-1β-stimulated SW982 cells, and RA-like fibroblast-like synoviocytes (FLS) were employed to investigate the effect and possible mechanism of osthole on arthritis in vivo and in vitro.
Results: 20 and 40 mg/kg osthole significantly alleviated collagen-induced arthritic symptoms based on histopathology and clinical arthritis scores, and improved erosion using HE staining. 20 and 40 mg/kg osthole decreased the level of IL-1β, TNF-α and IL-6 in rats and ameliorated oxidative stress in serum evaluated using ELISA kits. In addition, treatment with 50 and 100 μM osthole for 48 h inhibited 10 ng/ml IL-1β-stimulated proliferation and migration of SW982, and significantly inhibited the expression of matrix metalloproteinases, such as MMP-1, MMP-3 and MMP-13, as detected by western blot. 50 and 100 μM osthole also blocked the generation of IL-6 and TNF-α in IL-1β-stimulated SW982 cells. The NF-κB and MAPK pathways were also inhibited by osthole in IL-1β-treated SW982 cells.
Conclusion: These results collectively demonstrated that osthole improves collagen-induced arthritis in a rat model and IL-1β-treated SW982 cells through inhibiting inflammation and cellular stress in vivo and in vitro, and osthole might be a promising therapeutic agent for RA.
Keywords Osthole, Collagen-induced arthritis, Synovial fibroblasts, Inflammation, NF-κB
Address and Contact Information 1 Department of Osteology, YeDa Hospital, Taishan Road No. 11 Economic and Technological District of Yantai, Yantai 264006, China.
2 School of Pharmacy, Jiangsu university, Zhenjiang 212000, China.
* Corresponding author: doctoryangming1@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0088-y Volume 23 (2018)
Authors Mokgadi Violet Gwangwa, Anna Margaretha Joubert and Michelle Helen Visagie*
Abstract Tumourigenic tissue uses modified metabolic signalling pathways in order to support hyperproliferation and survival. Cancer-associated aerobic glycolysis resulting in lactic acid production was described nearly 100 years ago. Furthermore, increased reactive oxygen species (ROS) and lactate quantities increase metabolic, survival and proliferation signalling, resulting in increased tumourigenesis. In order to maintain redox balance, the cell possesses innate antioxidant defence systems such as superoxide dismutase, catalase and glutathione. Several stimuli including cells deprived of nutrients or failure of antioxidant systems result in oxidative stress and cell death induction. Among the cell death machinery is autophagy, a compensatory mechanism whereby energy is produced from damaged and/or redundant organelles and proteins, which prevents the accumulation of waste products, thereby maintaining homeostasis. Furthermore, autophagy is maintained by several pathways including phosphoinositol 3 kinases, the mitogen-activated protein kinase family, hypoxia-inducible factor, avian myelocytomatosis viral oncogene homolog and protein kinase receptor-like endoplasmic reticulum kinase. The persistent potential of cancer metabolism, redox regulation and the crosstalk with autophagy in scientific investigation pertains to its ability to uncover essential aspects of tumourigenic transformation. This may result in clinical translational possibilities to exploit tumourigenic oxidative status and autophagy to advance our capabilities to diagnose, monitor and treat cancer.
Keywords Warburg effect, Autophagy, Oxidative stress, Cancer
Address and Contact Information Department of Physiology, Faculty of Health Sciences, University of Pretoria, Private Bag X323, Arcadia 0007, South Africa
* Corresponding author: michelle.visagie@up.ac.za
Read full article at BMC

DOI: 10.1186/s11658-018-0085-1 Volume 23 (2018)
Authors Xisong Wang and Qing Song*
Abstract Background: Post-infarction cardiac injury is closely associated with cardiac remodeling and heart dysfunction. Mammalian STE20-like kinase 1 (Mst1), a regulator of cellular apoptosis, is involved in cardiac remodeling in post-infarction heart, but the mechanisms remain poorly defined. We aimed to explore the role of Mst1 in regulating chronic post-infarction cardiac injury, with a focus on mitochondrial homoeostasis.
Methods: Wild-type (WT) and Mst1-knockout mice were as the cardiac myocardial infarction model. Cardiac fibrosis, myocardial inflammation response, heart dysfunction and cardiomyocyte death were measured in vivo using immunohistochemistry, immunofluorescence, western blot, qPCR and TUNEL assays. Cardiomyocytes were isolated from WT and Mst1-knockout mice, and a chronic hypoxia model was used to induce damage. Mitochondrial function was determined via JC1 staining, ROS measurement, cyt-c leakage detection and mitochondrial apoptotic pathways analysis. Mitochondrial fission was observed using immunofluorescence. A pathway activator and inhibitor were applied to establish the signaling pathways involved in regulating mitochondrial homeostasis.
Results: Our study demonstrated that Mst1 expression was significantly upregulated in the heart post-infarction. Activated Mst1 induced cardiac fibrosis, an excessive inflammatory response, and cardiomyocyte death, whereas the genetic ablation of Mst1 protected the myocardium against chronic post-infarction injury. Function assays showed that upregulation of Mst1 activity contributed to JNK pathway activation, which led to Drp1 migration from the cytoplasm onto the surface of the mitochondria, indicative of mitochondrial fission activation. Excessive mitochondrial fission caused mitochondrial fragmentation, resulting in mitochondrial potential collapse, ROS overproduction, mitochondrial pro-apoptotic leakage into the cytoplasm, and the initiation of caspase-9-mediated mitochondrial apoptosis. By contrast, Mst1 deletion helped to maintain mitochondrial structure and function, sending pro-survival signals to the cardiomyocytes.
Conclusions: Our results identify Mst1 as a malefactor in the development of post-infarction cardiac injury and that it acts through the JNK-Drp1-mitochondrial fission pathway.
Keywords Cardiac post-infarction injury, Mst1, Mitochondrial fission, JNK-Drp1 pathway, Cardiomyocyte death
Address and Contact Information Department of Critical Care Medicine, the Chinese PLA General Hospital, Beijing, China
* Corresponding author: zhengdehuang198@hotmail.com
Read full article at BMC

DOI: 10.1186/s11658-018-0089-x Volume 23 (2018)
Authors Xiaowei Wang*, Haiying Jin, Shifeng Jiang and Yanlan Xu
Abstract Background: MicroRNAs (miRNAs) have physiological and pathophysiological functions that are involved in the regulation of cardiac fibrosis. This study aimed to investigate the effects of miR-495 on high glucose-induced cardiac fibrosis in human cardiac fibroblasts (CFs) and to establish the mechanism underlying these effects.
Methods: Human CFs were transfected with an miR-495 inhibitor or mimic and incubated with high glucose. The levels of NOD1 and miR-495 were then determined via quantitative RT-PCR. Pro-inflammatory cytokine levels, cell differentiation and extracellular matrix accumulation were respectively detected using ELISA, quantitative RT-PCR and western blot assays. The luciferase reporter assay, quantitative RT-PCR and western blot were used to explore whether NOD1 was a target of miR-495. The effects of miR-495 on the NF-κB and TGF-β1/Smad signaling pathways were also detected via western blot.
Results: Our results show that high glucose can significantly increase the expression of NOD1 in a time-dependent manner. Upregulation of miR-495 significantly alleviated the high glucose-induced increases in cell differentiation and collagen accumulation of CFs. Moreover, the bioinformatics analysis predicted that NOD1 was a potential target gene for miR-495. The luciferase reporter assay showed that miR-495 can directly target NOD1. The introduction of miR-495 could significantly inhibit the high glucose-activated NF-κB and TGF-β1/Smad signaling pathways.
Conclusion: Upregulation of miR-495 ameliorates the high glucose-induced inflammatory, cell differentiation and extracellular matrix accumulation of human CFs by modulating both the NF-κB and TGF-β1/Smad signaling pathways through downregulation of NOD1 expression. These results provide further evidence for the protective effect of miR-495 overexpression in cases of high glucose-induced cardiac fibrosis.
Keywords MicroRNA-495, Human cardiac fibroblasts, High glucose, Cardiac fibrosis, NOD1
Address and Contact Information Department of Geriatrics, Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University, Qingpu District, No.1158, Park East Road, Shanghai 201707, People’s Republic of China
* Corresponding author: wangxiaoweifudan@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0087-z Volume 23 (2018)
Authors Jing-jing Zhao, Xiao-Li Lou, Hong-wei Chen, Feng-ting Zhu and Yan-Qiang Hou*
Abstract The levels of decoy receptor 3 (DcR3), soluble urokinase type plasminogen activator receptor (suPAR) and procalcitonin (PCT) are significantly increased in sepsis. We investigated the diagnostic value of DcR3 combined with suPAR and PCT in sepsis. Patients with sepsis, non-infectious systemic inflammatory response comprehensive syndrome (SIRS) and healthy controls were recruited according to the diagnostic standard. We measured DcR3, suPAR, PCT, interleukin-6 (IL-6) and C-reactive protein (CRP), and the diagnostic value was evaluated by receiver operating characteristics (ROC) curves. In our analysis, serum DcR3, suPAR and PCT levels of the sepsis group were significantly higher than those of the SIRS and control groups. However, IL-6, CRP and WBC showed no significant difference between the SIRS group and the sepsis group. The serum DcR3 level was positively correlated with the serum suPAR level (r = 0.37, p = 0.0022) and PCT level (r = 0.37, p = 0.0021). Using DcR3, suPAR and PCT to distinguish SIRS from sepsis, the area under the curve (AUC) values were 0.892, 0.778 and 0.692. When DcR3, suPAR and PCT combined were used for diagnosis of sepsis, the AUC was 0.933, at a cut-off point of 0.342. This combination improved the sensitivity and specificity of diagnosis of sepsis, suggesting that use of the combination of three indexes enhanced the efficiency of sepsis diagnosis.
Keywords Sepsis, SIRS, DcR3, suPAR, PCT
Address and Contact Information Department of Central Laboratory, Songjiang Hospital Affiliated First People’s Hospital, Shanghai Jiao Tong University, Shanghai 201600, China
* Corresponding author: houyanqiang@aliyun.com
Equal contributors
Read full article at BMC

DOI: 10.1186/s11658-018-0091-3 Volume 23 (2018)
Authors Yanfen Zhang, Wenjing Zhang, Caijun Zha and Yanhong Liu*
Abstract Background: Patients with anti-β2GPI antibodies display significantly higher platelet activation/aggregation and vascular endothelial cell damage. The mechanism underlying the correlation between platelet activation, vascular endothelial cell dysfunctions and anti-β2GPI antibodies remains unknown.
Methods: In this study, we derived miR-96 and -26a from platelets activated by the anti- β2GPI/β2GPI complex and explored their role in modulating human umbilical vein endothelial cell (HUVEC) migration and tube formation.
Results: Anti-β2GPI/β2GPI complex induces the release of platelet-derived microparticles (p-MPs). The amounts of miR-96 and -26a in these p-MPs were also higher than for the control group. Co-incubation of HUVECs with p-MPs resulted in the transfer of miR-96 and -26a into HUVECs, where they inhibited migration and tube formation. The targeting role of these miRNAs was further validated by directly downregulating targeted selectin-P (SELP) and platelet-derived growth factor receptor alpha (PDGFRA) via luciferase activity assay.
Conclusion: Our study suggests that miR-96 and -26a in p-MPs can inhibit HUVEC behavior by targeting SELP and PDGFRA.
Keywords Platelet-derived microparticles, MicroRNA, Endothelial cells
Address and Contact Information Department of Laboratory Diagnosis, The Second Affliated Hospital of Harbin Medical University, Harbin, China
* Corresponding author: 460205012@qq.com
Read full article at BMC

DOI: 10.1186/s11658-018-0090-4 Volume 23 (2018)
Authors Aixia Sun1†, Hongwei Zhang1*†, Feng Pang1, Guifen Niu2, Jianzhong Chen3, Fei Chen3 and Jian Zhang4
Abstract Background: Although significant advances have been made in understanding the mechanisms of macrophage response to Staphylococcus aureus infection, the molecular details are still elusive. Identification of the essential genes and biological processes of macrophages that are specifically changed at different durations of S. aureus exposure is of great clinical significance.
Methods: We aimed to identify the significantly changed genes and biological processes of S. aureus-exposed macrophages. We systematically analyzed the macrophage gene expression profile GSE 13670 database with 8 h, 24 h or 48 h S. aureus infection. The results were further confirmed by western blot and quantitative polymerase chain reaction (qPCR) analyses.
Results: After 8 h of S. aureus infection, the expression of 624 genes was significantly changed. Six hundred thirteen differentially expressed genes (DEGs) were identified after 24 h of S. aureus infection. Two hundred fifty-three genes were significantly changed after 48 h of S. aureus infection. STAT1 was consistently up-regulated in these three treatments. TP53, JAK2, CEBPA, STAT3, MYC, CTNNB1 and PRKCA were only identified in the 8 h or 24 h S. aureus infection groups. CTNNB1 and PRKCA were for the first time identified as potential essential genes in S. aureus infection of macrophages. In the Gene Ontology (GO) term analysis, the defense response was shown to be the most significantly changed biological process among all processes; KEGG pathway analysis identified the JAK-STAT signaling pathway involved in early infection.
Conclusions: Our systematic analysis identified unique gene expression profiles and specifically changed biological processes of the macrophage response to different S. aureus exposure times.
Keywords Macrophage, Staphylococcus aureus, Bio-informatics
Address and Contact Information 1 Department of Clinical Laboratory, Liaocheng People’s Hospital, 67 West Dongchang Road, Liaocheng 252000, Shandong Province, People’s Republic of China.
2 Department of Endocrinology, Liaocheng People’s Hospital, 67 West Dongchang Road, Liaocheng 252000, Shandong Province, People’s Republic of China.
3 Department of Clinical Pharmacy, Liaocheng People’s Hospital, 67 West Dongchang Road, Liaocheng 252000, Shandong Province, People’s Republic of China.
4 Outpatient Vaccination Service, Center for Disease Control and Prevention of Liaocheng, Liaocheng 252000, Shandong Province, People’s Republic of China.
* Corresponding author: 13310659996@189.cn
Read full article at BMC

DOI: 10.1186/s11658-018-0077-1 Volume 23 (2018)
Authors Wenming Feng1*, Tao Xue2, Sanxiong Huang3, Qilin Shi4, Chengwu Tang1, Ge Cui4, Guanghui Yang5, Hui Gong2 and Huihui Guo2
Abstract Background: Hypoxia plays a critical role in many cancers. Hypoxia inducible factor-1α (HIF-1α) is an important mediator of the hypoxia response. It regulates the expression of various chemokines involved in tumor growth, angiogenesis and metastasis but the associated pathway needs further investigation.
Methods: The expression level of HIF-1α was determined in hepatocellular carcinoma (HCC) cells. The correlation of interleukin-8 (IL-8) and HIF-1α was assessed by knocking down HIF-1α. These cells were also used to assess its influence on HCC cell migration and invasion was checked. Pyrrolidinedithiocarbamate (PDTC), an inhibitor of NF-κB, was used to confirm the associated signaling pathway.
Results: HIF-1α was significantly expressed in HCC cells and found to promote HCC cell migration and invasion in an IL-8-dependent manner. NF-κB was confirmed to be involved in the process.
Conclusions: HIF-1α promotes HCC cell migration and invasion by modulating IL-8 via the NF-κB pathway.
Keywords Hepatocellular carcinoma, Hypoxia, HIF-1α, IL-8, NF-κB
Address and Contact Information 1 Department of Hepatobiliary Pancreatic Surgery, The First People’s Hospital of Huzhou, No. 158 Guangchanghou Road, Huzhou, Zhejiang Province 313000, People’s Republic of China.
2 Central Laboratory, The First People’s Hospital of Huzhou, No. 158 Guangchanghou Road, Huzhou, Zhejiang Province 313000, People’s Republic of China.
3 Department of General Surgery, The First People’s Hospital of Huzhou, No. 158 Guangchanghou Road, Huzhou, Zhejiang Province 313000, People’s Republic of China.
4 Department of Pathology, The First People’s Hospital of Huzhou, No. 158 Guangchanghou Road, Huzhou, Zhejiang Province 313000, People’s Republic of China.
5 Department of Pharmacy, The First People’s Hospital of Huzhou, No. 158 Guangchanghou Road, Huzhou, Zhejiang Province 313000, People’s Republic of China.
* Corresponding author: wenmingfeng2017@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0092-2 Volume 23 (2018)
Authors Hongxia Li1, Yi Cui2, Fuyan Li3, Wenqi Shi4, Wenjing Gao2, Xiao Wang2 and Qingshi Zeng2*
Abstract Background: Radiotherapy is among the commonly applied treatment options for glioma, which is one of the most common types of primary brain tumor. To evaluate the effect of radiotherapy noninvasively, it is vital for oncologists to monitor the effects of X-ray irradiation on glioma cells. Preliminary research had showed that PKC-ι expression correlates with tumor cell apoptosis induced by X-ray irradiation. It is also believed that the lactate-to-creatine (Lac/Cr) ratio can be used as a biomarker to evaluate apoptosis in glioma cells after X-ray irradiation. In this study, we evaluated the relationships between the Lac/Cr ratio, apoptotic rate, and protein kinase C iota (PKC-ι) expression in glioma cells.
Methods: Cells of the glioma cell lines C6 and U251 were randomly divided into 4 groups, with every group exposed to X-ray irradiation at 0, 1, 5, 10 and 15 Gy. Single cell gel electrophoresis (SCGE) was conducted to evaluate the DNA damage. Flow cytometry was performed to measure the cell cycle blockage and apoptotic rates. Western blot analysis was used to detect the phosphorylated PKC-ι (p-PKC-ι) level. 1H NMR spectroscopy was employed to determine the Lac/Cr ratio.
Results: The DNA damage increased in a radiation dose-dependent manner (p < 0.05). With the increase in X-ray irradiation, the apoptotic rate also increased (C6, p < 0.01; U251, p < 0.05), and the p-PKC-ι level decreased (C6, p < 0.01; U251, p < 0.05). The p-PKC-ι level negatively correlated with apoptosis, whereas the Lac/Cr ratio positively correlated with the p-PKC-ι level.
Conclusion: The Lac/Cr ratio decreases with an increase in X-ray irradiation and thus can be used as a biomarker to reflect the effects of X-ray irradiation in glioma cells.
Keywords DNA damage, P-PKC-ι, Lactate, Glioma, X-ray irradiation, Apoptosis
Address and Contact Information 1 Department of Radiology, the Second Hospital of Shandong University, Jinan, China.
2 Department of Radiology, Qilu Hospital of Shandong University, 107 Wenhuaxi Road, Jinan 250012, China.
3 Department of Radiology, Shandong Medical Imaging Research Institute, Jinan, China.
4 Department of Radiology, the Third Affiliated Hospital, Sun Yat- Sen University, Guangzhou, China.
* Corresponding author: zengqingshi@sina.com
Read full article at BMC

DOI: 10.1186/s11658-018-0094-0 Volume 23 (2018)
Authors Bingxia Shi*, Wei Yan, Guolin Liu and Yanjun Guo
Abstract Background: It has been reported that the expression of activating transcription factor 3 (ATF3) is closely associated with both microRNA (miRNA) processing and the progress of many cancers. Our study aimed to explore the interaction between ATF3 and miR-488 in tongue squamous cell carcinoma (TSCC).
Methods: Quantitative real-time PCR was performed to detect the levels of ATF3 and miR-488 in TSCC tissues and cell lines. Cell invasion and epithelial–mesenchymal transition (EMT) were assessed to determine the biological functions of miR-488 and ATF3 in TSCC cells. The mRNA and protein levels of ATF3 were measured using quantitative RT-PCR and western blotting. Luciferase assays were performed to validate ATF3 as an miR-488 target in TSCC cells.
Results: We found that the level of miR-488 significantly decreased and the expression of ATF3 significantly increased in TSCC tissues and cell lines. A low level of miR-488 was closely associated with increased expression of ATF3 in TSCC tissues. Introducing miR-488 significantly inhibited the invasion and EMT of TSCC cells, and knockdown of miR-488 promoted both processes. The bioinformatics analysis predicted that ATF3 is a potential target gene of miR-488. The luciferase reporter assay showed that miR-488 could directly target ATF3. ATF3 silencing had similar effects to miR-488 overexpression on TSCC cells. Overexpression of ATF3 in TSCC cells partially reversed the inhibitory effects of the miR-488 mimic.
Conclusion: miR-488 inhibited cell invasion and EMT of TSCC cells by directly downregulating ATF3 expression.
Keywords Tongue squamous carcinoma, MicroRNA-488, Activating transcription factor 3, Invasion, Epithelial–mesenchymal transition
Address and Contact Information Oral and Maxillofacial Surgery, Cangzhou Central Hospital, No. 16 Xinhua West Road, Cangzhou, Hebei 061000, People’s Republic of China
* Corresponding author: shibingxiacangzhou@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0095-z Volume 23 (2018)
Authors Jingjing Lu1, Feng Xu2, Yingna Zhang3, Hong Lu4 and Jiewen Zhang1*
Abstract Background: Mishandling of intracellular chloride (Cl) concentration ([Cl]i) in cerebrovascular smooth muscle cells is implicated in several pathological processes, including hyperplasia and remodeling. We investigated the effects of ClC-2-mediated Cl efflux on the proliferation of human brain vascular smooth muscle cells (HBVSMCs) induced by angiotensin II (AngII).
Methods: Cell proliferation and motility were determined using the CCK-8, bromodeoxyuridine staining, wound healing and invasion assays. ClC-2, PCNA, Ki67, survivin and cyclin D1 expression, and β-catenin and GSK-3β phosphorylation were examined using western blotting. Histological analyses were performed using hematoxylin and eosin staining and α-SMA staining.
Results: Our results showed that AngII-induced HBVSMC proliferation was accompanied by a decrease in [Cl]i and an increase in ClC-2 expression. Inhibition of ClC-2 by siRNA prevented AngII from inducing the efflux of Cl. AngII-induced HBVSMC proliferation, migration and invasion were significantly attenuated by ClC-2 downregulation. The inhibitory effects of ClC-2 knockout on HBVSMC proliferation and motility were associated with inactivation of the Wnt/β-catenin signaling pathway, as evidenced by inhibition of β-catenin phosphorylation and nuclear translocation, and decrease of GSK-3β phosphorylation and survivin and cyclin D1 expression. Recombinant Wnt3a treatment markedly reversed the effect of ClC-2 knockdown on HBVSMC viability. An in vivo study revealed that knockdown of ClC-2 with shRNA adenovirus ameliorated basilar artery remodeling by inhibiting Wnt/β-catenin signaling in AngII-treated mice.
Conclusion: This study demonstrates that blocking ClC-2-mediated Cl efflux inhibits AngII-induced cerebrovascular smooth muscle cell proliferation and migration by inhibiting the Wnt/β-catenin pathway. Our data indicate that downregulation of ClC-2 may be a viable strategy in the prevention of hyperplasia and remodeling of cerebrovascular smooth muscle cells.
Keywords Cerebrovascular smooth muscle cells, Proliferation, Angiotensin II, Chloride, ClC-2, Wnt/β-catenin signaling
Address and Contact Information 1 Department of Neurology, Henan People’s Hospital, No. 7 Wai-5 Road, Zhengzhou 450052, Henan Province, China.
2 Department of Urology, First Affiliated Hospital, Zhengzhou University, Zhengzhou, China.
3 Institute of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, China.
4 Department of Neurology, First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, Henan Province, China.
* Corresponding author: HZ_Sammy@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0097-x Volume 23 (2018)
Authors Junxia Zhang1,2, Baochun Yang1,2, Xiaocheng Wen1,2* and Guoqiang Sun2
Abstract Background: Bamei pigs are special germplasm resources in Qinghai. In the specific environment of the Qinghai plateau, local breeds, through long-term natural and artificial selection, have acquired a relatively stable population structure and genetic diversity. This study revealed Bamei pigs’ genetic diversity at the molecular level and the degree of introgression of commercial breeds into Bamei pigs.
Methods: The mitochondrial DNA D-loop region was amplified and sequenced using the ABI 3130 DNA sequencer. Sequences were collected, edited and aligned using the MEGA 5.0, DNASTAR and ClustalX 1.81 software. Haplotypes of all sequences and genetic diversity were analyzed by DnaSP 5.0 software.
Results and conclusions: We identified a total of 73 polymorphic sites in our 489 novel sequences, including 1 single variable site and 72 parsimony informative sites. Genetic diversity was analyzed in this study. The results showed that haplotype diversity, nucleotide diversity and the average number of nucleotide differences of Bamei pigs were lower than those of commercial pigs. Synthetic evaluation of genetic diversity through principal component analysis indicated that Bamei pigs also showed low genetic diversity. In summary, the results of sequence analysis showed that Bamei pigs were low in genetic diversity, and the introgression of commercial pigs into Bamei pigs was serious.
Keywords Mitochondrial DNA, Genetic variation, Bamei pig, Commercial pig
Address and Contact Information 1 State Key Laboratory of Plateau Ecology and Agriculture, Qinghai University, Xining 810016, Qinghai, China
2 College of Agriculture and Animal Husbandry, Qinghai University, Xining, Qinghai, China
* Corresponding author: 506944045@qq.com
Read full article at BMC

DOI: 10.1186/s11658-018-0093-1 Volume 23 (2018)
Authors Marta Pałka†, Aleksandra Tomczak†, Katarzyna Grabowska†, Magdalena Machowska, Katarzyna Piekarowicz, Dorota Rzepecka and Ryszard Rzepecki*
Abstract Lamin proteins are type V intermediate filament proteins (IFs) located inside the cell nucleus. They are evolutionarily conserved and have similar domain organization and properties to cytoplasmic IFs. Lamins provide a skeletal network for chromatin, the nuclear envelope, nuclear pore complexes and the entire nucleus. They are also responsible for proper connections between the karyoskeleton and structural elements in the cytoplasm: actin and the microtubule and cytoplasmic IF networks. Lamins affect transcription and splicing either directly or indirectly. Translocation of active genes into the close proximity of nuclear lamina is thought to result in their transcriptional silencing. Mutations in genes coding for lamins and interacting proteins in humans result in various genetic disorders, called laminopathies. Human genes coding for A-type lamin (LMNA) are the most frequently mutated. The resulting phenotypes include muscle, cardiac, neuronal, lipodystrophic and metabolic pathologies, early aging phenotypes, and combined complex phenotypes. The Drosophila melanogaster genome codes for lamin B-type (lamin Dm), lamin A-type (lamin C), and for LEM-domain proteins, BAF, LINC-complex proteins and all typical nuclear proteins. The fruit fly system is simpler than the vertebrate one since in flies there is only single lamin B-type and single lamin A-type protein, as opposed to the complex system of B- and A-type lamins in Danio, Xenopus and Mus musculus. This offers a unique opportunity to study laminopathies. Applying genetic tools based on Gal4 and in vitro nuclear assembly system to the fruit fly model may successfully advance knowledge of laminopathies. Here, we review studies of the laminopathies in the fly model system.
Keywords Lamins, Lamin Dm, Lamin C, Nuclear lamina, Nuclear envelope, Laminopathy, LMNA, Fruit fly
Address and Contact Information † Marta Pałka, Aleksandra Tomczak and Katarzyna Grabowska contributed equally to this work.
Laboratory of Nuclear Proteins, Faculty of Biotechnology, University of Wroclaw, Fryderyka Joliot-Curie 14a, 50-383 Wroclaw, Poland
* Corresponding author: rzepecki@ibmb.uni.wroc.pl
Read full article at BMC

DOI: 10.1186/s11658-018-0099-8 Volume 23 (2018)
Authors Shaoxian Liu1, Junyi Yue1, Wei Du2, Jian Han3 and Weidong Zhang4*
Abstract Background: Osteosarcoma (OS) is a common malignant tumor that predominantly occurs in adolescents. Its most common metastasis is to the lungs. As shown in our earlier study, lysosome-associated membrane glycoprotein 3 (LAMP3) is highly upregulated in metastatic OS. However, its role in the regulation of OS cell viability and apoptosis remains unknown.
Methods: We knocked down and overexpressed LAMP3 in OS cells and assessed the cell viability and apoptosis. Then, we investigated the expression of apoptosis-associated genes to identify the downstream gene(s) of LAMP3.
Results: Knockdown of LAMP3 significantly inhibited OS cell viability and promoted apoptosis. TP53, which is involved in the apoptosis pathway, was found to be highly upregulated after knockdown of LAMP3. Overexpression of LAMP3 significantly increased cell viability and abrogated apoptosis. Importantly, subsequent knockdown of TP53 partially suppressed the increased OS cell apoptosis induced by the inhibition of LAMP3, suggesting that TP53 is a key functional downstream gene of LAMP3.
Conclusions: Our findings suggest that LAMP3 promotes OS cell viability and survival by regulating TP53 expression.
Keywords Osteosarcoma, LAMP3, TP53
Address and Contact Information 1 Department of Bone Traumatology, Yantaishan Hospital, Yantai 264000, Shandong Province, People’s Republic of China.
2 Department of Spinal Research, Yantaishan Hospital, Yantai 264000, Shandong Province, People’s Republic of China.
3 Department of Bone Tumor, Yantaishan Hospital, Yantai264000Shandong Province, People’s Republic of China.
4 Department of Bone Traumatology, Yantai Hospital of Traditional Chinese Medicine, Yantai 264000, Shandong Province, People’s Republic of China.
* Corresponding author: 15605454815@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0096-y Volume 23 (2018)
Authors Nikolaos G. Nikitakis1,2†, Ioannis Gkouveris1†, Jaya Aseervatham1, Kelvin Barahona1 and Kalu U. E. Ogbureke1*
Abstract Background: Recent findings indicate that dentin sialophosphoprotein (DSPP) and matrix metalloproteinase (MMP) 20 interact in oral squamous cell carcinoma (OSCC). The objective of this study was to determine the effects of DSPP/MMP20 gene silencing on oral cancer stem cell (OCSC) markers.
Methods: The expression of well-established OCSC markers: ABCG2; ALDH1; CD133; CD44; BMI1; LGR4, and Podoplanin in DSPP/MMP20-silenced OSCC cell line, OSC2, and controls were assayed by western blot (WB), and flow cytometry techniques. The sensitivity of OSC2 cells to cisplatin following DSPP/MMP20 silencing was also determined.
Results: DSPP/MMP20 silencing resulted in downregulation of OCSC markers, more profoundly ABCG2 (84%) and CD44 (81%), following double silencing. Furthermore, while treatment of parent (pre-silenced) OSC2 cells with cisplatin resulted in upregulation of OCSC markers, DSPP/MMP20-silenced OSC2 cells similarly treated resulted in profound downregulation of OCSC markers (72 to 94% at 50 μM of cisplatin), and a marked reduction in the proportion of ABCG2 and ALDH1 positive cells (~ 1%).
Conclusions: We conclude that the downregulation of OCSC markers may signal a reduction in OCSC population following MMP20/DSPP silencing in OSCC cells, while also increasing their sensitivity to cisplatin. Thus, our findings suggest a potential role for DSPP and MMP20 in sustaining OCSC population in OSCCs, possibly, through mechanism(s) that alter OCSC sensitivity to treatment with chemotherapeutic agents such as cisplatin.
Keywords Cancer stem cells, Cancer stem cell markers, Oral Cancer, OSCC, MMP20, DSPP, Cisplatin
Address and Contact Information 1 Department of Diagnostic and Biomedical Sciences, University of Texas Health Sciences Center at Houston School of Dentistry, 7500 Cambridge Street, Houston, TX 77054, USA.
2 Department of Oral Pathology and Medicine, School of Dentistry, University of Athens, Athens, Greece.
* Corresponding author: Kalu.Ogbureke@uth.tmc.edu
† Nikolaos G. Nikitakis and Ioannis Gkouveris contributed equally to this work.
Read full article at BMC

DOI: 10.1186/s11658-018-0101-5 Volume 23 (2018)
Authors Ying Huang1,2†, Wuping Yang1,2†, Huihong Zeng2†, Chuan Hu2, Yaqiong Zhang2, Nanhua Ding2, Guangqin Fan1,3, Lijian Shao1,3* and Bohai Kuang2*
Abstract Upregulation of histone acetylation plays a critical role in the dysregulation of transcription. It alters the structure of chromatin, which leads to the onset of cancer. Histone deacetylase inhibitors may therefore be a promising way to limit cancer progression. In this study, we examined the effects of droxinostat on the growth of HT-29 colon cancer cells. Our results show that droxinostat effectively inhibited cell growth and colony-forming ability by inducing cellular apoptosis and ROS production in HT-29 cells. Notably, the apoptotic inhibitor Z-VAD-FMK significantly decreased the levels of cellular apoptosis and the antioxidant γ-tocotrienol (GT3) significantly decreased ROS production induced by droxinostat treatment. Z-VAD-FMK and GT3 also partially reversed the negative growth effects of droxinstat on HT-29 cells. GT3 treatment decreased cellular apoptosis and increased colony-forming ability upon droxinostat administration. Z-VAD-FMK treatment also partially decreased droxinostat-induced ROS production. Our findings suggest that the effects of droxinostat on colon cancer cells are mediated by the induction of oxidative stress and apoptotic cell death.
Keywords Droxinostat, HT-29 cells, Apoptosis, ROS
Address and Contact Information 1 Jiangxi provincial key laboratory of preventive medicine, Nanchang University, Nanchang 330006, China.
2 Medical School of Nanchang University, 461 Bayi Road, Nanchang 330006, Jiangxi, China.
3 School of Public Health, Nanchang University, Nanchang 330006, China.
* Corresponding author: lshao@ncu.edu.cn; kuangbohai@163.com
† Ying Huang, Wuping Yang and Huihong Zeng contributed equally to this work.
Read full article at BMC

DOI: 10.1186/s11658-018-0098-9 Volume 23 (2018)
Authors Jian-Peng Hu1†, Rong Zhang1†, Min Tang2, Yu-Lian Li2, Lin-Ting Xun2, Zhi-Zhou Shi1, Ying An2, Ting Li2 and Zheng-Ji Song2*
Abstract Background: We investigated the activity of loureirin B against liver fibrosis and the underlying molecular mechanisms.
Methods: Hepatic stellate cells (HSCs) from Sprague-Dawley rats were treated with different concentrations of loureirin B. We used the MTT assay to determine HSC proliferation, flow cytometry to analyze apoptosis, and western blot to determine the expressions of Bax, Bcl-2, Wnt1 and β-catenin. Real-time PCR was used to determine the expressions of Wnt1 and miR-148-3p.
Results: The MTT assay showed that loureirin B treatment significantly inhibited the proliferation of HSCs in time- and dose-dependent manners. Loureirin B significantly promoted the apoptosis of HSCs, increased the expression of Bax and decreased the Bcl-2 level. Western blot analysis showed that the expressions of Wnt1 and β-catenin were obviously lower in the loureirin B treatment group than in the control group. We also found that loureirin B could decrease the Wnt1 mRNA level and increase miR-148-3p expression. Knockdown of miR-148-3p using inhibitor could reverse the effects of loureirin B on the proliferation and apoptosis of HSCs and the expressions of Bax, Bcl-2, Wnt1 and β-catenin.
Conclusion: Our results suggest that loureirin B inhibited the proliferation and promoted the apoptosis of HSCs, and suppressed the Wnt/β-catenin signaling pathway via regulation of miR-148-3p.
Keywords Loureirin B, Liver fibrosis, Wnt1, miR-148-3p, Hepatic stellate cells
Address and Contact Information 1 Medical School, Kunming University of Science and Technology, Kunming 650500, Yunnan province, China.
2 Department of Gastroenterology, the First People’s Hospital of Yunnan province, the Affiliated Hospital of Kunming University of Science and Technology, Kunming 650032, Yunnan province, China.
* Corresponding author: song4715@163.com
† Jian-Peng Hu and Rong Zhang contributed equally to this work.
Read full article at BMC

DOI: 10.1186/s11658-018-0103-3 Volume 23 (2018)
Authors Liqun Chen*†, Lingjuan Wu†, Linyan Zhu and Yiyi Zhao
Abstract The nuclear receptor RXRα (retinoid X receptor-α) is a transcription factor that regulates the expression of multiple genes. Its non-genomic function is largely related to its structure, polymeric forms and modification. Previous research revealed that some non-genomic activity of RXRα occurs via formation of heterodimers with Nur77. RXRα–Nur77 heterodimers translocate from the nucleus to the mitochondria in response to certain apoptotic stimuli and this activity correlates with cell apoptosis. More recent studies revealed a significant role for truncated RXRα (tRXRα), which interacts with the p85α subunit of the PI3K/AKT signaling pathway, leading to enhanced activation of AKT and promoting cell growth in vitro and in animals. We recently reported on a series of NSAID sulindac analogs that can bind to tRXRα through a unique binding mechanism. We also identified one analog, K-80003, which can inhibit cancer cell growth by inducing tRXRα to form a tetramer, thus disrupting p85α–tRXRα interaction. This review analyzes the non-genomic effects of RXRα in normal and tumor cells, and discusses the functional differences based on RXRα protein structure (structure source: the RCSB Protein Data Bank).
Keywords Nuclear receptor, RXRα, Non-genomic action, Modification, Structure, RCSB protein data Bank
Address and Contact Information College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China
* Corresponding author: lqchen@fzu.edu.cn
† Liqun Chen and Lingjuan Wu contributed equally to this work.
Read full article at BMC

DOI: 10.1186/s11658-018-0102-4 Volume 23 (2018)
Authors Wei Wu1*, Li Nie2, Li Zhang3 and Yan Li1*
Abstract Background: Oncogenic Notch1 is known to activate the NF-κB pathway in T cell acute lymphoblastic leukemia (T-ALL) and to up-regulate the transcription of Asb2α, a specificity factor for an E3 ubiquitin ligase complex that plays an important role in hematopoietic differentiation. Therefore, we hypothesize that Notch1 might regulate the NF-κB pathway through Asb2α.
Methods: The study involved down-regulation of Notch1 in T-ALL cell lines (CCRF-CEM cells and MOLT-4 cells) through treatment with gamma-secretase inhibitor (GSI) as well as the modulation of Asb2 in CCRF-CEM cells and MOLT-4 cells through transduction with lentivirus carrying Asb2 or Asb2-shRNA. Experiments using real-time PCR, western blot and co-immunoprecipitation were performed to evaluate the expression levels of related genes. Cell proliferation and apoptosis were measured while the expression of Asb2 was enhanced or inhibited.
Results: Here, we demonstrated for the first time that Notch1 can activate the transcription of Asb2α, which then stimulates activation of NF-κB in T-ALL cells. Asb2α exerts its effects by inducing degradation and dissociation of IκBα from NF-κB in T-ALL cells. Moreover, specific suppression of Asb2α expression can promote apoptosis and inhibit proliferation of T-ALL cells.
Conclusion: Notch1 modulates the NF-κB pathway through Asb2α, indicating that Asb2α inhibition is a promising option for targeted therapy against T-ALL.
Keywords Notch1, NF-κB, Asb2, T cell acute lymphoblastic leukemia
Address and Contact Information 1 Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan 430060, People’s Republic of China.
2 Department of Geriatrics, Renmin Hospital of Wuhan University, Wuhan 430060, People’s Republic of China.
3 Department of Hematology, Renmin Hospital of Wuhan University, Wuhan 430060, People’s Republic of China.
* Corresponding author: wuwei_vivi2005@126.com; yanlitf@yahoo.com.cn
Read full article at BMC

DOI: 10.1186/s11658-018-0107-z Volume 23 (2018)
Authors Guangzhe Yu1, Qian Liu2, Xuening Dong2, Kaihong Tang2, Bohui Li2, Chunmei Liu2, Wenzheng Zhang2, Yiduo Wang2 and Yingyu Jin2*
Abstract Background: Inflammation is an important pathogenic component of endotoxemia-induced acute kidney injury (AKI), finally resulting in renal failure. Diacerein is an interleukin-1β (IL-1β) inhibitor used for osteoarthritis treatment by exerting anti-inflammatory effects. This study aims to investigate the effects of diacerein on endotoxemia-induced AKI.
Methods: Male C57BL/6 mice were intraperitoneally injected with lipopolysaccharide (LPS, 10 mg/kg) for 24 h prior to diacerein treatment (15 mg/kg/day) for another 48 h. Mice were examined by histological, molecular and biochemical approaches.
Results: LPS administration showed a time-dependent increase of IL-1β expression and secretion in kidney tissues. Diacerein treatment normalized urine volume and osmolarity, reduced blood urea nitrogen (BUN), fractional excretion of sodium (FENa), serum creatinine and osmolarity, and protected renal function in an endotoxemic AKI mice model. In the histopathologic study, diacerein also improved renal tubular damage such as necrosis of the tubular segment. Moreover, diacerein inhibited LPS-induced increase of inflammatory cytokines, such as IL-1β, tumor necrosis factor-α, monocyte chemoattractant protein-1 and nitric oxide synthase 2. In addition, LPS administration markedly decreased aquaporin 1 (AQP1), AQP2, AQP3, Na,K-ATPase α1, apical type 3 Na/H exchanger and Na-K-2Cl cotransporter expression in the kidney, which was reversed by diacerein treatment. We also found that diacerein or IL-1β inhibition prevented the secretion of inflammatory cytokines and the decrease of AQP and sodium transporter expression induced by LPS in HK-2 cells.
Conclusion: Our study demonstrates for the first time that diacerein improves renal function efficiently in endotoxemic AKI mice by suppressing inflammation and altering tubular water and sodium handing. These results suggest that diacerein may be a novel therapeutic agent for the treatment of endotoxemic AKI.
Keywords Endotoxemia, Acute kidney injury, Inflammation, Aquaporins, Sodium transporters, Diacerein
Address and Contact Information 1 Department of Emergency Surgery, The 1st Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang Province, China.
2 Department of Laboratory Diagnosis, The 1st Affiliated Hospital of Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin 150001, Heilongjiang Province, People’s Republic of China.
* Corresponding author: yingyu_jin@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0104-2 Volume 23 (2018)
Authors Fengrui Wu1,2*, Qingqing Wu1,2, Dengkun Li1,2, Yuan Zhang1,2, Rong Wang1,2, Yong Liu1,2 and Wenyong Li1,2*
Abstract Background: The transcription factor Oct4 plays a pivotal role in the pre- implantation development of the mouse embryo. DNA methyltransferase 1 (Dnmt1) maintains the changes in DNA methylation during mammalian early embryonic development. Little is known of the role of Oct4 in DNA methylation in mice. In this study, Kunming white mice were used as an animal model to reveal any correlation between DNA methylation and Oct4 during mammalian embryonic development.
Results: The expressions of Dnmt1 and Oct4 were initially studied using real-time PCR. They exhibited different patterns during the pre-implantation stage. Moreover, by using a promoter assay and ChIP analysis, we found that the transcriptional activities of Dnmt1 in mouse NIH/3 T3 cells and CCE cells were regulated by Oct4 through direct binding to the − 554 to − 294 fragment of the upstream regulation element of Dnmt1. The downregulation of Dnmt1 expression and enzyme activity by mouse Oct4 were further confirmed by transfecting Oct4 siRNA into mouse CCE cells.
Conclusion: Our results indicate that Oct4 is involved in DNA methylation through the regulation of Dnmt1 transcription, especially during the early stages of mouse pre-implantation embryo development.
Keywords Dnmt1, Oct4, Transcription regulation, Kunming white mouse
Address and Contact Information 1 Anhui Province Key Laboratory of Embryo Development and Reproductive Regulation, Fuyang Normal University, Fuyang, China.
2 Anhui Province Key Laboratory of Environmental Hormone and Reproduction, Fuyang Normal University, Fuyang, China.
* Corresponding author: fengruiwu@126.com; liwenyong@aliyun.com
Read full article at BMC

DOI: 10.1186/s11658-018-0106-0 Volume 23 (2018)
Authors Xin Ni1,2†, Xin-zhi Li3†, Zhi-ru Fan1,2†, Ai Wang1,2, Hai-chao Zhang1,2, Liang Zhang1,2*, Li LI1,2, Jun-qiang Si1,2 and Ke-tao Ma1,2*
Abstract Background: Imbalances in circulating T lymphocytes play critical roles in the pathogenesis of hypertension-mediated inflammation. Connexins (Cxs) in immune cells are involved in the maintenance of homeostasis of T lymphocytes. However, the association between Cxs in peripheral blood T lymphocytes and hypertension-mediated inflammation remains unknown. This study was designed to investigate the role of Cxs in T lymphocytes in hypertension-mediated inflammation in spontaneously hypertensive rats (SHRs).
Methods: The systolic blood pressure (SBP) in Wistar-Kyoto (WKY) rats and SHRs was monitored using the tail-cuff method. The serum cytokine level was determined using ELISA. The proportions of different T-lymphocyte subtypes in the peripheral blood, the expressions of Cx40/Cx43 in the T-cell subtypes, and the gap junctional intracellular communication (GJIC) of peripheral blood lymphocytes were measured using flow cytometry (FC). The accumulations of Cx40/Cx43 at the plasma membrane and/or in the cytoplasm were determined using immunofluorescence staining. The in vitro mRNA levels of cytokines and GJIC in the peripheral blood lymphocytes were respectively examined using real-time PCR and FC after treatment with Gap27 and/or concanavalin A (Con A).
Results: The percentage of CD4+ T cells and the CD4+/CD8+ ratio were high, and the accumulation or expressions of Cx40/Cx43 in the peripheral blood lymphocytes in SHRs were higher than in those of WKY rats. The percentage of CD8+ and CD4+ CD25+ T cells was lower in SHRs. The serum levels of IL-2, IL-4 and IL-6 from SHRs were higher than those from WKY rats, and the serum levels of IL-2 and IL-6 positively correlated with the expression of Cx40/Cx43 in the peripheral blood T lymphocytes from SHRs. The peripheral blood lymphocytes of SHRs exhibited enhanced GJIC. Cx43-based channel inhibition, which was mediated by Gap27, remarkably reduced GJIC in lymphocytes, and suppressed IL-2 and IL-6 mRNA expressions in Con A stimulated peripheral blood lymphocytes.
Conclusions: Our data suggest that Cxs may be involved in the regulation of T-lymphocyte homeostasis and the production of cytokines. A clear association was found between alterations in Cxs expression or in Cx43-based GJIC and hypertension-mediated inflammation.
Keywords Hypertension-mediated inflammation, T lymphocytes, Connexins, Spontaneously hypertensive rats
Address and Contact Information 1,2 Department of Physiology of the Medical College of Shihezi University, 59 North 2nd Road, Shihezi, Xinjiang 832002, People’s Republic of China
3 Department of Pathophysiology of the Medical College of Shihezi University, 59 North 2nd Road, Shihezi, Xinjiang 832002, People’s Republic of China
* Corresponding author: maketao@hotmail.com; zhangliang_0622@163.com
† Xin Ni, Xin-zhi Li and Zhi-ru Fan contributed equally to this work.
Read full article at BMC

DOI: 10.1186/s11658-018-0105-1 Volume 23 (2018)
Authors Zhi-Qiang Tian1, Hong Jiang2* and Zhi-Bing Lu2
Abstract Background: MicroRNAs play important roles in regulation of the cardiovascular system. The purpose of this study was to investigate microRNA-320 (miR-320) expression in myocardial ischemia-reperfusion (I/R) injury and the roles of miR-320 in cardiomyocyte apoptosis by targeting AKIP1 (A kinase interacting protein 1).
Methods: The level of miR-320 was detected using quantitative real-time polymerase chain reaction (qRT-PCR), and cardiomyocyte apoptosis was detected via terminal dUTP nick end-labeling assay. Cardiomyocyte apoptosis and the mitochondrial membrane potential were evaluated via flow cytometry. Bioinformatics tools were used to identify the target gene of miR-320. The expression levels of AKIP1 mRNA and protein were detected via qRT-PCR and Western blot, respectively.
Results: Both the level of miR-320 and the rate of cardiomyocyte apoptosis were substantially higher in the I/R group and H9c2 cells subjected to H/R than in the corresponding controls. Overexpression of miR-320 significantly promoted cardiomyocyte apoptosis and increased the loss of the mitochondrial membrane potential, whereas downregulation of miR-320 had an opposite effect. Luciferase reporter assay showed that miR-320 directly targets AKIP1. Moreover, knock down and overexpression of AKIP1 had similar effects on the H9c2 cells subjected to H/R.
Conclusions: miR-320 plays an important role in regulating cardiomyocyte apoptosis induced by I/R injury by targeting AKIP1 and inducing the mitochondrial apoptotic pathway.
Keywords microRNA-320, Ischemia–reperfusion, AKIP1, Cardiomyocyte apoptosis
Address and Contact Information 1 Department of Cardiology, Inner Mongolia People’s Hospital, Hohhot 010017, People’s Republic of China.
2 Department of Cardiology, Remin Hospital of Wuhan University, 238 Jiefang Road, Wuhan 430060, People’s Republic of China.
* Corresponding author: hongj0505@126.com; 13848182619@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0109-x Volume 23 (2018)
Authors Mohammad Reza Abbaszadegan1, Anali Riahi2, Mohammad Mahdi Forghanifard3 and Meysam Moghbeli4*
Abstract Background: Esophageal squamous cell carcinoma (ESCC) is the most common histological type of esophageal cancer, with a poor prognosis. Deregulation of WNT and NOTCH signaling pathways is important in ESCC progression, which can be due to either malfunction of their components or crosstalk with other pathways. Therefore, identification of new crosstalk between such pathways may be effective to introduce new strategies for targeted therapy of cancer. A correlation study was performed to assess the probable interaction between growth factor receptors and WNT/NOTCH pathways via the epidermal growth factor receptor (EGFR) and Musashi1 (MSI1), respectively.
Methods: Levels of MSI1/EGFR mRNA expression in tumor tissues from 48 ESCC patients were compared to their corresponding normal tissues using real-time polymerase chain reaction.
Results: There was a significant correlation between EGFR and MSI1 expression (p = 0.05). Moreover, there was a significant correlation between EGFR/MSI1 expression and grade of tumor differentiation (p = 0.02).
Conclusion: This study confirms a direct correlation between MSI1 and EGFR and may support the important role of MSI1 in activation of EGFR through NOTCH/WNT pathways in ESCC.
Keywords Growth factor, WNT, NOTCH, Early stage, Survival
Address and Contact Information 1 Medical Genetics Research Center, Faculty of Medical Sciences, Mashhad University of Medical Sciences, Mashhad, Iran.
2 Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
3 Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran.
4 Department of Modern Sciences and Technologies, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
* Corresponding author: Meysam_moghbeli@yahoo.com
Read full article at BMC

DOI: 10.1186/s11658-018-0108-y Volume 23 (2018)
Authors Lifan Zhuang1, Jianhua Tian1, Xinzhi Zhang1, Hong Wang1 and Chenghui Huang1,2*
Abstract Background: Lnc-DC is a specific group of long non-coding (Lnc) RNAs in dendritic cells (DCs). Its function has been previously studied, and includes roles in dendritic cell differentiation and the progression of some diseases. In this study, we observed the critical role of Lnc-DC in regulating the differentiation, growth, and apoptosis of dendritic cells.
Methods: We first isolated peripheral blood mononuclear cells to culture and induce into DCs, which were then co-cultured with hepatitis B virus (HBV)-secreting HepG2. 2.15 cells for the detection of changes in Lnc-DC. The expression levels of TLR9, p-STAT3, and SOCS3 were tested with qPCR and western blot. MTT assays were used to analyze the cell proliferation, cell cycle, and apoptosis. We used ELISA to test the expression of TNF-α, IL-1β, IL-6, IL-12p40, and IFN-γ.
Results: Co-culture with HBV-secreting HepG2.2.15 cells increased the level of Lnc-DC and activated TLR9/STAT3 signaling. The HBV DNA level (IU/ml) was positively correlated with levels of Lnc-DC and TLR9, further demonstrating that Lnc-DC was associated with the immune response of HBV. Lnc-DC was shown to regulate TLR9/ STAT3 signaling in dendritic cells. More interestingly, the regulation of Lnc-DC controlled the immune response by reducing the concentration of secreted TNF-α, IL-6, IL-12, and IFN-γ, as well as increasing the IL-1β concentration in dendritic cells.
Conclusion: Lnc-DC is important in regulating the growth, apoptosis, and immune response of dendritic cells mediated by TLR9/STAT3 signaling, and was also activated by HBV. This study provides a previously unidentified mechanism underlying the immune response in dendritic cells.
Keywords Dendritic cell, Lnc-DC, TLR9/STAT3, HBV
Address and Contact Information 1 Department of Infectious Disease, the Affiliated Shenzhen Baoan Hospital of Southern Medical University, Shenzhen 518101, China
2 Department of Infectious Disease, Shenzhen Baoan District People’s Hospital, No. 118, Xin’an Street, Long Jing er Raod, Shenzhen 518101, China
* Corresponding author: huangchenghuisci@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0110-4 Volume 23 (2018)
Authors Yanxia Wang1, Hui Xu1, Baoan Zhu2, Zhenling Qiu3*† and Zaijun Lin4*†
Abstract Background: Tumor microenvironment, in particular the stroma, plays an important role in breast cancer cell invasion and metastasis. Investigation of the molecular characteristics of breast cancer stroma may reveal targets for future study.
Methods: The transcriptome profiles of breast cancer stroma and normal breast stroma were compared to identify differentially expressed genes (DEGs). The method was analysis of GSE26910 and GSE10797 datasets. Common DEGs were identified and then analyses of enriched pathways and hub genes were performed.
Results: A total of 146 DEGs were common to GSE26910 and GSE10797. The enriched pathways were associated with “extracellular matrix (ECM) organization”, “ECM-receptor interaction” and “focal adhesion”. Network analysis identified six key genes, including JUN, FOS, ATF3, STAT1, COL1A1 and FN1. Notably, COL1A1 and FN1 were identified for the first time as cancer stromal key genes associated with breast cancer invasion and metastasis. Oncome analysis showed that the high expression levels of COL1A1 and FN1 correlated to an advanced stage of breast cancer and poor clinical outcomes.
Conclusions: We found that several conserved tumor stromal genes might regulate breast cancer invasion through ECM remodeling. The clinical outcome analyses of COL1A1 and FN1 suggest these two genes are promising targets for future studies.
Keywords Differentially expressed genes, COL1A1, FN1, Breast cancer
Address and Contact Information 1 Department of Emergency Medicine, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Center for Specialty Strategy Research of Shanghai Jiao Tong University China Hospital, Shanghai 200000, People’s Republic of China.
2 Department of Biochemistry, Luohe Medical College, Luohe 462002, Henan Province, People’s Republic of China.
3 Shandong Yantai Laiyang Center Hospital, 111 Changshan Road, Laiyang 265200, Shandong Province, China.
4 The Orthopedic Department of Shanghai Hospital of Traditional Chinese Medicine, 274 Zhijiang Middle Road, Jing’an District, Shanghai 200000, People’s Republic of China.
* Corresponding author: qiuzhenling@hotmai.com; lin-zaijun@163.com
† Zhenling Qiu and Zaijun Lin contributed equally to this work.
Read full article at BMC

DOI: 10.1186/s11658-018-0111-3 Volume 23 (2018)
Authors Rafal Bartoszewski1* and Aleksander F. Sikorski2*
Abstract Recent developments in high-throughput genotyping technologies have revealed the existence of several new classes of RNA that do not encode proteins but serve other cellular roles. To date, these non-coding RNAs (ncRNAs) have been shown to modulate both gene expression and genome remodeling, thus contributing to the control of both normal and disease-related cellular processes. The attraction of this research topic can be seen in the increasing number of submissions on ncRNAs to molecular biology journals, including Cellular Molecular Biology Letters (CMBL). As researchers attempt to deepen the understanding of the role of ncRNAs in cell biology, it is worth discussing the broader importance of this research.
Keywords ncRNAs, microRNAs, lncRNAs, Regulation of gene expression
Address and Contact Information 1 Department of Biology and Pharmaceutical Botany, Medical University of Gdansk, Gdansk, Poland
2 Department of Cytobiochemistry, Faculty of Biotechnology, University of Wroclaw, Wroclaw, Poland
* Correspondin author: rafalbar@gumed.edu.pl; aleksander.sikorski@uwr.edu.pl
Read full article at BMC

DOI: 10.1186/s11658-018-0100-6 Volume 23 (2018)
Authors Maoyun Fei1, Jianming Guan2, Tao Xue3, Lianjin Qin1, Chengwu Tang4, Ge Cui1, Yao Wang1, Hui Gong3 and Wenming Feng4*
Abstract Background: Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third most common cause of cancer-related death worldwide. The 5-year survival rate remains low despite considerable research into treatments of HCC, including surgery, radiotherapy and chemotherapy. Many mechanisms within HCC still require investigation, including the influence of hypoxia, which has a crucial role in many cancers and is associated with metastasis. Hypoxia inducible factor-1α (HIF-1α) is known to regulate the expression of many chemokines, including interleukin-8 (IL-8), which is associated with tumor metastasis. Although many studies have reported that HIF-1α is associated with HCC migration and invasion, the underlying mechanisms remain unknown.
Methods: The expression level of HIF-1α was determined in HCC cells. The correlation of IL-8 and HIF-1α expressions was assessed via knockdown of HIF-1α. HCC cells were also used to assess the influence of HIF-1α on HCC cell migration and invasion. LY294002, an inhibitor of the Akt pathway, was used to confirm the associated signaling pathways.
Results: We observed a significant attenuation of cell migration and invasion after silencing of HIF-1α. Exogenously expressing IL-8 restored migration and invasion. Akt was found to be involved in this process.
Conclusion: Hypoxia promotes HCC cell migration and invasion through the HIF-1α–IL-8–Akt axis.
Keywords Hepatocarcinoma, Hypoxia, HIF-1α, IL-8, Akt pathway
Address and Contact Information 1Department of General Surgery, The First People’s Hospital of Huzhou, No.158 Guangchanghou RoadZhejiang Province 313000 Huzhou, People’s Republic of China.
2 Department of Ultrasound, The First People’s Hospital of Huzhou, No.158 Guangchanghou RoadZhejiang Province 313000 Huzhou, People’s Republic of China.
3 Central Laboratory, The First People’s Hospital of Huzhou, No.158 Guangchanghou RoadZhejiang Province 313000 Huzhou, People’s Republic of China.
4 Department of Hepatobiliary Pancreatic Surgery, The First People’s Hospital of Huzhou, No.158 Guangchanghou RoadZhejiang Province 313000 Huzhou, People’s Republic of China.
* Corresponding author: wenmingfeng2018@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0114-0 Volume 23 (2018)
Authors Zhong-Shi Xu1*, Feng Dai2, Ji Chen1, Meng Lv1, Ji-Wu Cheng1, Xiao-Ming Zhang1 and Bo-Wen Lin1
Abstract Background: Evidence has shown that endogenous H2S plays an important role in the physiological and pathophysiological processes of many organs. The study aimed to explore whether exogenous H2S has a potential therapeutic effect on a rat ovariectomy-induced model of osteoporosis.
Methods: The OVX osteoporosis model was established in female Sprague-Dawley rats by full bilateral ovariectomy. The rats were randomly divided into four groups, with the two experimental groups receiving an intraperitoneal injection of GYY4137 or sodium alendronate. The level of H2S in the plasma was determined and common laboratory indicators to diagnose osteoporosis, such as alkaline phosphatase (ALP) activity and the levels of osteocalcin (OCN), calcitonin, parathyroid hormone and leptin were measured. The bone mineral density (BMD) of the 4th and 5th lumbar vertebrae was measured using dual-energy X-ray absorptiometry. The maximum stress of femoral fracture was obtained through a three-point bending test of the femur.
Results: The OVX osteoporosis model was successfully established. GYY4137 was injected to increase the level of H2S in the plasma in one group, designated OVX-GYY during the observation period (p < 0.05). At 12 weeks, the BMD value of the fourth lumbar vertebra in the OVX-GYY group had increased (p < 0.05). The BMD femur value in the OVX-vehicle group had decreased (p < 0.05). Bilateral ovariectomy leads to biochemical disorders related to bone metabolism and hormone levels in rat plasma (all p < 0.05). Ovariectomy also reduced blood calcium, blood phosphate and calcitonin, and increased parathyroid hormone and leptin. The opposite results were obtained for the groups with alendronate sodium or GYY4137 treatment (all p < 0.05).
Conclusions: Through the slow release of H2S, GYY4137 did an excellent job of simulating endogenous neuroendocrine gaseous signaling molecules. Exogenous H2S had a regulatory effect on osteoporosis in ovariectomized rats, showing potential value for the treatment of human postmenopausal osteoporosis.
Keywords H2S, Osteoporosis, Ovariectomized, Bone metabolism, Bone mineral density
Address and Contact Information 1 Department of Orthopedics, Second Clinical Medical College of Jinan University (Shenzhen People’s Hospital), Dongmen North Road 1017, Luohu District, Shenzhen 518020, China.
2 Department of Radiology, Second Clinical Medical College of Jinan University (Shenzhen People’s Hospital), Shenzhen 518020, China.
* Corresponding author: 13509670866@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0112-2 Volume 23 (2018)
Authors Grey W Fortenbery1, Brinda Sarathy2, Kristen R Carraway2 and Kyle D Mansfield2*
Abstract Background: Tissue ischemia can arise in response to numerous physiologic and pathologic conditions. The cellular response to decreased perfusion, most notably a decrease in glucose and oxygen, is important for cellular survival. In response to oxygen deprivation or hypoxia, one of the key response elements is hypoxia inducible factor (HIF) and a key protein induced by hypoxia is vascular endothelial growth factor (VEGF). Under hypoxia, we and others have reported an increase in the half-life of VEGF and other hypoxia related mRNAs including MYC and CYR61; however, the mediator of this response has yet to be identified. For this study, we sought to determine if HIF-mediated transcriptional activity is involved in the mRNA stabilization induced by hypoxia.
Methods: HEK293T or C6 cells were cultured in either normoxic or hypoxic (1% oxygen) conditions in the presence of 1 g/L glucose for all experiments. Pharmacological treatments were used to mimic hypoxia (desferroxamine, dimethyloxaloglutamate, CoCl2), inhibit mitochondrial respiration (rotenone, myxothiazol), scavenge reactive oxygen species (ROS; ebselen), or generate mitochondrial ROS (antimycin A). siRNAs were used to knock down components of the HIF transcriptional apparatus. mRNA half-life was determined via actinomycin D decay and real time PCR and western blotting was used to determine mRNA and protein levels respectively.
Results: Treatment of HEK293T or C6 cells with hypoxic mimetics, desferroxamine, dimethyloxaloglutamate, or CoCl2 showed similar induction of HIF compared to hypoxia treatment, however, in contrast to hypoxia, the mimetics caused no significant increase in VEGF, MYC or CYR61 mRNA half-life. Knockdown of HIF-alpha or ARNT via siRNA also had no effect on hypoxic mRNA stabilization. Interestingly, treatment of HEK293T cells with the mitochondrial inhibitors rotenone and myxothiazol, or the glutathione peroxidase mimetic ebselen did prevent the hypoxic stabilization of VEGF, MYC, and CYR61, suggesting a role for mtROS in the process. Additionally, treatment with antimycin A, which has been shown to generate mtROS, was able to drive the normoxic stabilization of these mRNAs.
Conclusion: Overall these data suggest that hypoxic mRNA stabilization is independent of HIF transcriptional activity but requires mtROS.
Keywords Hypoxia, Hypoglycemia, HIF, Mitochondrial reactive oxygen species, mRNA stability
Address and Contact Information 1 Brody School of Medicine, East Carolina University, Greenville, NC 27834, USA.
2 Biochemistry and Molecular Biology, Brody School of Medicine, East Carolina University, Greenville, NC 27834, USA.
* Corresponding author: mansfieldk@ecu.edu
Read full article at BMC

DOI: 10.1186/s11658-018-0113-1 Volume 23 (2018)
Authors Toru Miyanaga1,4*, Yoshimichi Ueda2, Aiko Miyanaga3, Mikio Yagishita1 and Naoko Hama1
Abstract To date, studies on mesenchymal tissue stem cells (MSCs) in the perichondrium have focused on in vitro analysis, and the dynamics of cartilage regeneration from the perichondrium in vivo remain largely unknown. We have attempted to apply cell and tissue engineering methodology for ear reconstruction using cultured chondrocytes. We hypothesized that by inducing angiogenesis with basic fibroblast growth factor (bFGF), MSCs or cartilage precursor cells would proliferate and differentiate into cartilage in vivo and that the regenerated cartilage would maintain its morphology over an extended period. As a result of a single administration of bFGF to the perichondrium, cartilage tissue formed and proliferated while maintaining its morphology for at least 3 months. By day 3 post bFGF treatment, inflammatory cells, primarily comprising mononuclear cells, migrated to the perichondrial region, and the proliferation of matrix metalloproteinase 1 positive cells peaked. During week 1, the perichondrium thickened and proliferation of vascular endothelial cells was noted, along with an increase in the number of CD44-positive and CD90-positive cartilage MSCs/progenitor cells. Neocartilage was formed after 2 weeks, and hypertrophied mature cartilage was formed and maintained after 3 months. Proliferation of the perichondrium and cartilage was bFGF concentration-dependent and was inhibited by neutralizing antibodies. Angiogenesis induction by bFGF was blocked by the administration of an angiogenesis inhibitor, preventing perichondrium proliferation and neocartilage formation. These results suggested that angiogenesis may be important for the induction and differentiation of MSCs/cartilage precursor cells in vivo, and that morphological changes, once occurring, are maintained.
Keywords Angiogenesis, Basic fibroblast growth factor, Differentiation, Elastic cartilage, In vivo model, Mesenchymal stem cell, Progenitor cell, Proliferation
Address and Contact Information 1 Department of Plastic and Reconstructive Surgery, Kanazawa Medical University, 1-1 Daigaku, Uchinada-machi, Kahoku-gun, Ishikawa 9200293, Japan.
2 Department of Pathology, Kanazawa Medical University, 1-1 Daigaku, Uchinada-machi, Kahoku-gun, Ishikawa 9200293, Japan.
3 Department of Nursing, Kanazawa Medical University, 1-1 Daigaku, Uchinada-machi, Kahoku-gun, Ishikawa 9200293, Japan.
4 Kanazawa Medical University Hospital, 1-1 Daigaku, Uchinada-machi, Kahoku-gun, Ishikawa 9200293, Japan.
* Corresponding author: miyanaga@kanazawa-med.ac.jp
Read full article at BMC

DOI: 10.1186/s11658-018-0115-z Volume 23 (2018)
Authors Yonghua Xu1†, Xiangmin Wang2†, Surong Jiang2, Chen Men2, Di Xu2, Yan Guo2 and Jun Wu2*
Abstract Background: Microcystins are waterborne environmental toxins that induce oxidative stress and cause injuries in the heart. On the other hand, many physiological processes, including antioxidant defense, are under precise control by the mammalian circadian clock.
Results: In the present study, we evaluated the effect of microcystin-LR (MC-LR) on the rhythmic expression patterns of circadian and antioxidant genes in rat cardiomyocytes using the serum shock technique. We found that a non-toxic dose (10 μm) of MC-LR decreased the amplitudes of rhythmic patterns of clock genes, while it increased the expression levels of antioxidant genes.
Conclusions: Our results indicate an influence of MC-LR on the circadian clock system and clock-controlled antioxidant genes, which will shed some light on the explanation of heart toxicity induced by MC-LR from the viewpoint of chronobiology.
Keywords Circadian clock, Antioxidant defense, Microcystin-LR, Heart
Address and Contact Information 1 Department of General Surgery, The Fourth Affiliated Hospital of Nantong University, Yancheng 224006, China.
2 Department of Geriatric Cardiology, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Rd, Nanjing 210029, China.

* Corresponding author: wujun9989@njmu.edu.cn
Yonghua Xu and Xiangmin Wangcontributed equally to this work.
Read full article at BMC

DOI: 10.1186/s11658-018-0116-y Volume 23 (2018)
Authors Shadan Hajalirezay Yazdi1, Mahdi Paryan2 and Samira Mohammadi-Yeganeh3,4,5*
Abstract Background: Breast cancer is the most prevalent cancer among women, and AXL and MET are the key genes in the PI3K/AKT/mTOR pathway as critical elements in proliferation and invasion of cancer cells. MicroRNAs (miRNAs) are small non-coding RNAs regulating the expression of genes.
Methods: Bioinformatic approaches were used to find a miRNA that simultaneously targets both AXL and MET 3′-UTRs. The expression of target miRNA was evaluated in triple-negative (MDA-MB-231) and HER2-overexpressing (SK-BR-3) breast cancer cell lines as well as normal breast cells, MCF-10A, using quantitative real-time PCR. Then, the miRNA was overexpressed in normal and cancer cell lines using a lentiviral vector system. Afterwards, effects of overexpressed miRNA on the expression of AXL and MET genes were evaluated using quantitative real-time PCR.
Results: By applying bioinformatic software and programs, miRNAs that target the 3′-UTR of both AXL and MET mRNAs were determined, and according to the scores, miR-34a was selected for further analyses. The expression level of miR-34a in MDA-MB-231 and SK-BR-3 was lower than that of MCF-10A. Furthermore, AXL and MET expression in SK-BR-3 and MDA-MB-231 was lower and higher, respectively, than that of MCF-10A. After miR-34a overexpression, MET and AXL were downregulated in MDA-MB-231. In addition, MET was downregulated in SK-BR-3, while AXL was upregulated in this cell line.
Conclusions: These findings may indicate that miR-34a is an oncogenic miRNA, downregulated in the distinct breast cancer subtypes. It also targets MET and AXL 3′-UTRs in triple-negative breast cancer. Therefore, it can be considered as a therapeutic target in this type of breast cancer.
Keywords Breast cancer, miRNA-34a, AXL, MET, Real-time PCR
Address and Contact Information 1 Department of Cellular and Molecular Biology, Faculty of Advanced Sciences & Technology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
2 Department of Research and Development, Production and Research Complex, Pasteur Institute of Iran, Tehran, Iran.
3 Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
4 Department of Biotechnology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
5 Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
* Corresponding author: s.mohammadiyeganeh@sbmu.ac.ir
Read full article at BMC

DOI: 10.1186/s11658-018-0117-x Volume 23 (2018)
Authors Yafeng Liang1,2, Nengli Yang3, Guoquan Pan2, Bingxin Jin2, Shufen Wang2 and Wei Ji1*
Abstract Background: Pulmonary inflammation and endothelial barrier permeability increase in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) induced by pro-inflammatory cytokines and matrix metalloproteinases (MMPs). However, the relationship between pro-inflammatory cytokines and MMPs in ALI/ARDS remains poorly understood.
Methods: A lipopolysaccharide (LPS)-induced ALI rat model was established through intratracheal instillation. The wet/dry ratios of lung tissues were measured, and bronchoalveolar lavage fluid (BALF) was collected to test protein concentrations, total cell/macrophage numbers, and pro-inflammatory cytokine levels. LPS-treated alveolar macrophages were utilized in in vitro experiments. The expression and secretion of MMPs were respectively detected using quantitative PCR, Western blotting and ELISA assays.
Results: The levels of IL-33 and MMP2/9 in BALF increased in all the ALI rats with severe lung injury. LPS-induced IL-33 autocrine upregulated the expression of MMP2 and MMP9 through activating STAT3. Neutralizing IL-33 in culture medium with specific antibodies suppressed the expression and secretion of MMP2 and MMP9 in LPS-treated alveolar macrophages. Consistently, eliminating IL-33 decreased the levels of MMP2 and MMP9 in BALF and alleviated lung injury in ALI rats.
Conclusion: The IL-33/STAT3/MMP2/9 regulatory pathway is activated in alveolar macrophages during acute lung injury, which may exacerbate the pulmonary inflammation.
Keywords Acute lung injury (ALI), IL-33, MMP2, MMP9, STAT3
Address and Contact Information 1 Department of Pediatric Pulmonology, Children’s Hospital of Soochow University, No 303, Jingde Road, Suzhou 215003, China.
2 Department of Pediatric Intensive Care Unit, The Second Affiliated Hospital & Yuying Children’s Hospital, Wenzhou Medical University, Wenzhou 325000, China.
3 Department of Anesthesiology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
* Corresponding author: szdxjiwei@163.com
Read full article at BMC

DOI: 10.1186/s11658-018-0120-2 Volume 23 (2018)
Authors Xing’an Zhang1,2, Lanfang Zhang3, Xiaoyao Tan1, Ying Lin4, Xinsheng Han1, Huadong Wang1, Huawei Ming1, Qiujiang Li2, Kang Liu2* and Gang Feng2*
Abstract Oral cancer remains a deadly disease worldwide. Lymph node metastasis and invasion is one of the causes of death from oral cancer. Elucidating the mechanism of oral cancer lymph node metastasis and identifying critical regulatory genes are important for the treatment of this disease. This study aimed to identify differentially expressed genes (gene signature) and pathways that contribute to oral cancer metastasis to lymph nodes. The GSE70604-associated study compared gene profiles in lymph nodes with metastasis of oral cancer to those of normal lymph nodes. The GSE2280-associated study compared gene profiles in primary tumor of oral cancer with lymph node metastasis to those in tumors without lymph node metastasis. There are 28 common differentially expressed genes (DEGs) showing consistent changes in both datasets in overlapping analysis. GO biological process and KEGG pathway analysis of these 28 DEGs identified the gene signature CCND1, JUN and SPP1, which are categorized as key regulatory genes involved in the focal adhesion pathway. Silencing expression of CCND1, JUN and SPP1 in the human oral cancer cell line OECM-1 confirmed that those genes play essential roles in oral cancer cell invasion. Analysis of clinical samples of oral cancer found a strong correlation of these genes with short survival, especially JUN expression associated with metastasis. Our study identified a unique gene signature – CCND1, JUN and SPP1 – which may be involved in oral cancer lymph node metastasis.
Keywords Oral cancer, Metastasis, CCND1, JUN, SPP1
Address and Contact Information 1 Department of Stomatology, Nanchong Central Hospital, The Second Clinical Medical College of North Sichuan Medical College, Nanchong, Sichuan 637000, People’s Republic of China.
2 Institute of Tissue Engineering and Stem Cells, Nanchong Central Hospital, The Second Clinical Medical College of North Sichuan Medical College, No. 95, People’s south Road, Shunqing District, Nanchong, Sichuan 637000, People’s Republic of China.
3 Department of Burn and Plastic Surgery, Nanchong Central Hospital, The Second Clinical Medical College of North Sichuan Medical College, Nanchong, Sichuan 637000, People’s Republic of China.
4 Department of Science and Education, Nanchong Central Hospital, The Second Clinical Medical College of North Sichuan Medical College, Nanchong, Sichuan 637000, People’s Republic of China.
* Corresponding author: liukangnsmc@163.com; fenggangncch@163.com
Read full article at BMC