Lin F.,First Peoples Hospital of Taizhou |
Zhang C.-H.,First Peoples Hospital of Taizhou |
Zhou Z.-J.,First Peoples Hospital of Taizhou |
Liu F.,Shanghai University |
And 2 more authors.
Medical Journal of Chinese People's Liberation Army | Year: 2014
Objective To evaluate the effect of hepatocellular carcinoma suppressor gene-1 (HCCS1) on the cancer stemlike phenotype, and invasive and metastatic ability of liver tumor cells. Methods Cells of human liver tumor cell line MHCC-97H were divided into three groups: normal control group, HCCS1 group and Ad-LacZ group. Cells in normal control group received no specific treatment, and cells in HCCS1 group and Ad-LacZ group were infected with Ad-HCCS1 and Ad-LacZ respectively, with infestation rate of 100:1. Real-time PCR and Western blotting were used to assess the mRNA and protein expression of HCCS1. FACS was used to identify the proportion of CD133 and CD90 positive cells. ELISA was employed to measure the levels of VEGF in the supernatant. Western blotting was used to detect the E-cadherin and fibronectin expression. Transwell assay was conducted to test the invasive and metastatic ability for the tumor cells. Results The mRNA and protein expression of HCCS1 in the HCCS1 group were higher than those in normal control group and Ad-LacZ group (P<0.01). Compared with normal control and Ad- LacZ group, the proportion of CD133 and CD90 positive cells, the content of VEGF in the supernatant, and the number of cells infiltrating Transwell membrane significantly decreased, while the expression of E-cadherin and fibronectin significantly increased in the HCCS1 group (P<0.01). Conclusion HCCS1 could inhibit cancer stem-like phenotype, and invasive and metastatic ability of MHCC-97H cells.
Yang H.,Peking University |
Yang H.,Sun Yat Sen University |
Gao F.,Huazhong University of Science and Technology |
Li X.,First Peoples Hospital of Taizhou |
And 3 more authors.
Connective Tissue Research | Year: 2015
Tumor necrosis factor-α (TNF-α) has been shown to have a catabolic effect on intervertebral disc degeneration (IVDD), including increasing MMP3 expression and subsequent extracellular matrix (ECM) degradation. In contrast, transforming growth factor-β1 (TGF-β1) has an anabolic effect on nucleus pulposus (NP) cells. However, the anti-catabolic effect of TGF-β1 under inflammatory condition is unknown. The aim of this study was to demonstrate whether TGF-β1 can reverse TNF-α-induced MMP3 increase in NP cells and to further investigate the underlying mechanisms. The transcriptional activity, gene expression, and protein levels of MMP3 were measured by luciferase reporter assay, qRT-PCR and western blot, respectively. TNF-α increased MMP3 expression in rat NP cells time and dose dependently. TGF-β1 could abolish TNF-α-mediated up-regulation of collagen I and MMP3 expression, and down-regulate aggrecan and collagen II expression. The ERK1/2 signaling pathway was activated after exposure to TGF-β1. Treatment with ERK1/2 inhibitors (PD98059 and U0126) abolished the antagonistic effect of TGF-β1 on TNF-α mediated catabolic responses. These findings provide novel evidence supporting the anti-catabolic role of TGF-β1 in IVDD, which is important for the potential clinical application of TGF-β1 in disc degenerative disorders. © 2015 Taylor & Francis Group, LLC.
Chen X.-R.,First Peoples Hospital of Taizhou |
Wang J.-W.,First Peoples Hospital of Taizhou |
Li X.,Zhejiang Province Hospital |
Zhang H.,First Peoples Hospital of Taizhou |
Ye Z.-Y.,Zhejiang Province Hospital
World Journal of Gastroenterology | Year: 2010
AIM: To investigate the relation between gastric cancer and microsatellite instability (MSI), loss of heterozygosity (LOH) and promoter region methylation. METHODS: Fifty primary gastric carcinoma specimens were collected from patients with no family history of cancer. In addition, normal tissues were also collected from patients as controls. DNA was extracted by polymerase chain reaction for single-strand conformation polymorphism, bisulfite DNA sequencing, and methylation-specific band analysis. RESULTS: The positive rate for MSI and LOH in gastric carcinoma was 16% and 20%, respectively. According to the tumor, node and metastasis staging system, the LOH frequency was higher in gastric carcinoma at stages III and IV than in gastric carcinoma at stages I and II (P = 0.01), which was also significantly correlated with lymph node metastasis and clinicopathological characteristics of gastric carcinoma. Methylation of bone morphogenetic protein 3 (BMP3) gene promoter was detected in 64.44% of gastric carcinoma tissue samples. However, no statistical significance was observed between promoter region methylation and carcinoma differentiation. Interestingly, the BMP3 gene methylation rate was 71.05% and 28.58%, respectively, in MSI positive and negative cases (P = 0.031), suggesting that BMP3 genetic instability and promoter methylation are initiated during gastric carcinogenesis. LOH was detected mostly in the late stages of gastric carcinoma, indicating that gastric carcinoma at late stages has a higher infiltration and a poorer prognosis. CONCLUSION: Promotor region methylation of the BMP3 gene may cause gastric carcinoma in Chinese people. © 2010 Baishideng.
Yang H.,Sun Yat Sen University |
Liu H.,Sun Yat Sen University |
Li X.,First Peoples Hospital of Taizhou |
Pan H.,Sun Yat Sen University |
And 3 more authors.
Connective Tissue Research | Year: 2015
Syndecan-4 is emerging as an important player in cell interaction with the extracellular environment and has been shown to be involved in the progression of intervertebral disc degeneration. However, the mechanism of syndecan-4 regulation by TNF-α and the role of TGF-β1 in regulating syndecan-4 expression remain poorly understood in nucleus pulposus (NP) cells. The aim of this study was to investigate these mechanisms. We exposed NP cells to TNF-α and the gene, protein expression, and promoter activity levels of syndecan-4 were measured by qPCR, western blotting, and the luciferase reporter assay, respectively. The activation of the MAPK and NF-κB pathways was detected using western blot analysis. Syndecan-4 expression in rat NP cells was increased by TNF-α, but this was neither time nor dose dependent in response to TNF-α. ERK1/2, JNK, and NF-κB pathways were activated following TNF-α treatment. Treatment with ERK1/2 and NF-κB inhibitors decreased the up-regulation of syndecan-4 by TNF-α. However, JNK inhibition showed no effect on syndecan-4 expression induced by TNF-α. TNF-α mediated up-regulation of syndecan-4 was antagonized by TGF-β1. This study provided evidence for the differential regulation by MAPK and NF-κB pathways in the over-expression of syndecan-4 promoted by TNF-α in NP cells. Our results demonstrate that TGF-β1 exerts anabolic effects on intervertebral discs by inhibiting the expression of syndecan-4. © 2015 Informa Healthcare USA, Inc.
Huang H.,Fujian Medical University |
Huang H.,Taizhou Municipal Hospital |
Zhu Y.,Fujian Medical University |
Li S.,First Peoples Hospital of Taizhou
Molecular Medicine Reports | Year: 2015
There is currently a requirement for effective treatment strategies for human hepatocellular carcinoma (HCC), a leading cause of cancer-associated mortality. MicroRNA-122 (miR-122), a repressor of the endogenous apoptosis regulator Bcl-w, is frequently downregulated in HCC. Thus, it is hypothesized that the activation of miR-122 may induce selective hepatocellular apoptosis via caspase activation in a model of HCC. In the present study, an miR-122 mimic transfection was performed in HepG2 cells, and used to investigate the role and therapeutic potential of miR-122 in the regulation of HCC-derived cell lines. The apoptotic rates of HepG2 cells were significantly increased following miR-122 mimic transfection. Reverse transcription-polymerase chain reaction analysis revealed that Bcl-w mRNA was significantly reduced, while the mRNA levels of caspase-9 and caspase-3 were markedly increased. The immunocytochemistry results supported the mRNA trends. Collectively, the present results suggest that endogenous miR-122 contributes to HepG2 apoptosis and that transfection of mimic miR-122 normalizes apoptotic levels in a model of HCC.
Wang J.,Fudan University |
Yang Y.,Fudan University |
Guo S.,Fudan University |
Chen Y.,Fudan University |
And 13 more authors.
Genes and Immunity | Year: 2013
Ankylosing spondylitis (AS) is a chronic inflammatory disease with complex genetic traits. Multiple sequence variations have been associated with AS, but explained only a proportion of heritability. The studies herein aimed to explore potential associations between genomic copy number (CN) variation (CNV) and AS in Han Chinese. Five AS patients were examined with the high-density comparative genomic hybridization microarrays in the first screen test for AS-associated CNVs. A total of 533 AS patients and 792 unrelated controls were examined in confirmation studies with the AccuCopy assays. A significant association was observed between the CNV of HLA-DQA1 and that of AS. Compared with controls, AS patients showed an aberrant CN, and a significantly increased number of patients had more than two copies of HLA-DQA1. Therefore, the CNV of HLA-DQA1 may have an important role in susceptibility to AS in the Han Chinese population. © 2013 Macmillan Publishers Limited.
Zhou H.-P.,Municipal Hospital of Anqing |
Mou C.-H.,First Peoples Hospital of Taizhou |
Wu J.,First Peoples Hospital of Taizhou
Fudan University Journal of Medical Sciences | Year: 2011
Objective To observe the focal neurological metabolic abnormalities and its relationship with the prognosis in patients with primary brain stem injury (PBSI) by proton magnetic resonance spectroscopy 0 H-MRS). Methods Thirty-six PBSI patients were enrolled as study group underwent 'H-MRS between 3 hours and 16 days after injury. Twelve volunteers were enrolled as control group. Linear regression analysis was used to assess the correlation between ' H-MRS value and clinical index and Glasgow outcome score (GOS) 6 months after injury. Results Compared with controls, ventral pontine NAA/Cr and NAA/Cho values decreased, and Cho/Cr value increased (P<0.05) in PBSZ patient,and the difference got more obvious as injury aggravated. Patients with poor prognosis had lower NAA/Cr and NAA/Cho values than those with favourable prognosis (P<0.05). In study group,NAA and GOS was positively correlated (r=0.79, P<0.05) , while Cho/NAA and GOS was negatively correlated (r=-0.65,P < 0.05). Conclusions Analysis of 'H-MRS in PBSI patients at the early stage was useful in evaluating the severity of injury and prognosis.
Guan S.,Huazhong University of Science and Technology |
Wang Z.,First Peoples Hospital of Taizhou |
Xin F.,Huazhong University of Science and Technology |
Xin H.,First Peoples Hospital of Taizhou
Molecular Medicine Reports | Year: 2014
Vascular calcification significantly affects the health of the elderly. Increasing evidence proved that vascular calcification is an actively regulated osteogenic process. The osteochondrocytic differentiation of mesenchymal stem cells (MSCs) is a significant step of osteogenic processes. The Wnt pathways has been identified as contributing to the regulation of osteogenic mineralization during development and disease. However, it remains unknown whether these MSCs in the vascular calcification differentiate into normal vascular smooth muscle cells (VSMCs) in vivo in order to treat damaged vascular tissue or into calcified VSMCs to aggravate calcification correlated to the Wnt pathways. Thus, it is necessary to analyze the mechanisms of MSC differentiation in detail. In the present study a cell-cell co-culturing in vitro system was used to observe MSCs that directly interact with normal or calcified VSMCs during calcification and to investigate the gene expression of the Wnt pathways during the process. Direct co-cultures were established by seeding two different cell types, VSMCs or calcified VSMCs, or a mixture of both at ratios of 5,000:5,000 cells/1.7 cm2 onto either gelatin-coated 1.7-cm2 chamber slides for immunohistochemical analysis or gelatin-coated 75-cm2 tissue culture flasks for protein or RNA isolation. Osteoblastic differentiation was evaluated by examining the cell morphology and assessing the activity of alkaline phosphatase in the cell lysates by alkaline phosphatase staining. Additionally, the mRNA expression levels of the genes encoding for proteins involved in the Wnt signaling proteins, Wnt5A, LRP6, Ror2, c-Jun-N-terminal kinase and β-catenin, were assessed in each group. The present study demonstrated that Wnts are expressed in the progress of differentiation of MSCs during calcification. MSCs can differentiate into different cell phenotypes when there is direct cell-cell contact with VSMCs or calcified VSMCs, and the Wnt5a/Ror2 signaling pathway may be associated with the determination of differentiation of MSCs in this process.
PubMed | First Peoples Hospital of Taizhou
Type: Journal Article | Journal: World journal of gastroenterology | Year: 2010
To investigate the relation between gastric cancer and microsatellite instability (MSI), loss of heterozygosity (LOH) and promoter region methylation.Fifty primary gastric carcinoma specimens were collected from patients with no family history of cancer. In addition, normal tissues were also collected from patients as controls. DNA was extracted by polymerase chain reaction for single-strand conformation polymorphism, bisulfite DNA sequencing, and methylation-specific band analysis.The positive rate for MSI and LOH in gastric carcinoma was 16% and 20%, respectively. According to the tumor, node and metastasis staging system, the LOH frequency was higher in gastric carcinoma at stages III and IV than in gastric carcinoma at stages I and II (P = 0.01), which was also significantly correlated with lymph node metastasis and clinico- pathological characteristics of gastric carcinoma. Methylation of bone morphogenetic protein 3 (BMP3) gene promoter was detected in 64.44% of gastric carcinoma tissue samples. However, no statistical significance was observed between promoter region methylation and carcinoma differentiation. Interestingly, the BMP3 gene methylation rate was 71.05% and 28.58%, respectively, in MSI positive and negative cases (P = 0.031), suggesting that BMP3 genetic instability and promoter methylation are initiated during gastric carcinogenesis. LOH was detected mostly in the late stages of gastric carcinoma, indicating that gastric carcinoma at late stages has a higher infiltration and a poorer prognosis.Promotor region methylation of the BMP3 gene may cause gastric carcinoma in Chinese people.