【摘要】 目的 探索miR-203a-3p对人肝癌细胞系增殖和凋亡的分子调节机制。方法 RT-qPCR检测临床肝癌(HCC)组织及肝癌细胞系中miR-203a-3p的表达；建立miR-203a-3p过表达的HepG2细胞模型；MTT法检测细胞增殖；流式细胞测量术检测细胞凋亡；平板集落形成实验检测集落形成；细胞划痕实验检测细胞迁移；表达谱芯片检测mRNA水平；ELISA检测转化生长因子-β1(TGF-β1)的分泌；Western blot检测TGF-β1表达。结果 miR-203a-3p在HCC组织中低表达(P<0.05),与肿瘤分期呈负相关(P<0.05);HepG2细胞中过表达miR-203a-3p可显著降低细胞增殖速率(P<0.05)、迁移能力(P<0.05)、克隆形成能力(P<0.05),促进细胞凋亡(P<0.05);TGF-β1表达降低(P<0.05),而TGFβR1表达升高(P<0.05);细胞培养上清中TGF-β1含量降低(P<0.05);细胞内TGF-β1的蛋白降低(P<0.05);加入TGF-β1重组蛋白可减轻miR-203a-3p导致的增殖降低和迁移降低(P<0.05)。结论 miR-203a-3p抑制人肝细胞癌细胞系增殖，促凋亡，并且抑制TGF-β1蛋白的表达。更多还原

【Abstract】 Objective To explore the molecular mechanism of regulation on the proliferation and apoptosis in hepatocellular carcinoma(HCC) cells by miR-203a-3p. Methods RT-qPCR was used to detect the expression of miR-203a-3p in the sample of liver cancer tissues and liver cancer cell lines. The HepG2 cell model expressing miR-203a-3p was established. Cell proliferation was observed by MTT method and the apoptosis of cells was detected by flow cytometry. Plate colony formation assay was used to detect clone formation and cell scratch assay was used to detect cell migration. Expression profile chip was applied in the detection of mRNA and ELISA was used to detect the secretion of transforming growth factor-β1(TGF-β1). TGF-β1 protein expression was examined by Western blod technology. Results miR-203a-3p was weekly expressed in HCC tissues(P<0.001)and was negatively correlated with tumor stage(P<0.05). Over-expression of miR-203a-3p in HepG2 cells significantly reduced cell proliferation(P<0.05), migration(P<0.05), clone formation(P<0.05) and promoted cell apoptosis(P<0.05); TGF-β1 expression decreased(P<0.05) while the expression of TGFβR1 increased(P<0.05); Thecontent of TGF-β1 in the cell culture supernatant decreased(P<0.05). The protein of TGF-β1 in cells decreased(P<0.05) and TGF-β1 recombinant protein reversed the miR-203a-3p induced inhibition of proliferation as well as migration(P<0.05). Conclusions miR-203a-3p inhibits proliferation of human hepatocellular carcinoma cell lines, promotes apoptosis and inhibits the expression of TGF-β1 protein.更多还原