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目的观察靶向封闭survivin基因对肝癌细胞株HepG2增殖和凋亡的影响,探讨survivin基因在肝癌的发生、发展中的作用。方法设计1对survivin-siRNA,体外转录制备siRNA。应用脂质体转染技术将survivin-siRNA转染肝癌细胞株HepG2,应用有荧光标记的非特异性小分子siRNA检测转染效率;RT-PCR检测survivin-mRNA水平;SABC免疫组化染色技术检测survivin蛋白表达情况;MTT法检测细胞生长情况;流式细胞仪分析细胞凋亡。结果转染survivin-siRNA可使HepG2细胞survivin-mRNA水平下降53.8%,并可使survivin蛋白表达明显减少;转染survivin-siRNA对细胞增殖及细胞凋亡无明显影响(P>0.05)。结论抑制survivin表达对肝癌细胞株HePG2凋亡及增殖无明显影响。提示survivin在肝癌细胞增殖与调亡调控中所起的作用可能并非是关键性的。
Objective To observe the effect of the targeted survivin gene on the proliferation and apoptosis of hepatocellular carcinoma cell line HepG2 and to explore the role of survivin gene in the occurrence and development of hepatocellular carcinoma. Methods A pair of survivin-siRNA was designed to transcribe siRNA in vitro. Survivin-siRNA was transfected into HepG2 cells by lipofectamine 2000. Fluorescently labeled siRNA was used to detect transfection efficiency. Survivin-mRNA level was detected by RT-PCR. Survivin expression was detected by immunohistochemical staining (SABC) Protein expression; MTT assay cell growth; apoptosis was analyzed by flow cytometry. Results Survivin-siRNA transfected HepG2 cells decreased the survivin-mRNA level by 53.8% and significantly decreased the expression of survivin protein. Survivin-siRNA transfected survivin-siRNA had no effect on cell proliferation and apoptosis (P> 0.05). Conclusion Inhibition of survivin expression has no significant effect on the apoptosis and proliferation of HepG2 cells. This suggests that the role of survivin in the regulation of proliferation and apoptosis of hepatoma cells may not be crucial.