【摘要】 目的探讨微小RNA 204(miR-204)在肝细胞癌中的表达、临床意义及可能分子机制。方法收集手术切除的60例肝细胞癌及对应癌旁肝组织,实时定量PCR(qRT-PCR)检测miR-204在肝癌及癌旁组织中的表达,免疫组织化学染色检测miR-204下游潜在靶点Bcl-2与组蛋白脱乙酰酶1(Sirt1)的表达;用人工合成的miR-204模拟物转染人SMMC-7721肝癌细胞,MTT法及流式细胞术检测SMMC-7721细胞的增殖、凋亡的情况,qRT-PCR、Western blot法分别检测Bcl-2与Sirt1的mRNA和蛋白表达。结果 miR-204在肝癌组织中表达水平显著低于对应癌旁组织;肝癌组织中miR-204低表达与肿瘤大小、肿瘤个数、肿瘤TNM分期显著相关;miR-204低表达组Bcl-2与Sirt1蛋白表达显著高于miR-204高表达组,相关性分析结果显示肝癌组织中miR-204与Bcl-2、Sirt1蛋白表达呈显著负相关;miR-204可显著抑制SMMC-7721细胞的增殖并促进其凋亡,并下调Bcl-2与Sirt1的mRNA与蛋白表达水平。结论 miR-204在肝癌组织中表达下调并与肝癌恶性临床病理特征有关,miR-204抑制肝癌细胞增殖、促进其凋亡的作用可能与下调Bcl-2和Sirt1表达有关。更多还原

【Abstract】 Objective To investigate the level of microRNA-204( miR-204) in human hepatocellular carcinoma( HCC)and its effect on the potential mechanism of tumorgenesis. Methods Real-time quantitative PCR( qRT-PCR) was applied to detect the expression of miR-204 in HCC( n = 60) and the corresponding tumor-adjacent normal tissues. The expressions of Bcl-2 and Sirt1 were measured by immunohistochemistry( IHC). The artificial miR-204 was transiently transfected into human SMMC-7721 cells in vitro. The proliferation and apoptosis of the cells were detected by MTT assay and flow cytometry,respectively. The expression levels of Bcl-2 and Sirt1 mRNA and protein were determined by qRT-PCR and Western blotting,respectively. Results The expression level of miR-204 in HCC tissues was significantly lower than that in the adjacent normal tissues,and it was associated with tumor size,number of tumors and advanced TNM stage. The expressions of Bcl-2 and Sirt1 in the lower miR-204 level group were both higher than those in the higher miR-204 level group. Correlation analysis showed that miR-204 expression was negatively correlated with Bcl-2 and Sirt1 protein expression levels. Over-expressed miR-204 in SMMC-7721 cells suppressed cell proliferation and promoted cell apoptosis,and down-regulated mRNA and protein expressions of both Bcl-2 and Sirt1. Conclusion The expression of miR-204 in HCC tissues was significantly lower than that in tumor-adjacent normal tissues. miR-204 could inhibit HCC cell proliferation and induce apoptosis by down-regulating the expressions of Bcl-2 and Sirt1.更多还原