mcl-1特异性siRNA对HepG2细胞增殖及凋亡的影响

Effects of Mcl-1 Specific SiRNA on Proliferation and Apoptosis of HepG2 Cells

目的:肝癌分子靶向治疗是目前研究的热点,肝癌相关基因mcl-1在肝癌增殖及凋亡中的作用尚不明确,本研究拟探讨mcl-1特异性siRNA对体外培养肝癌细胞HepG2增殖及凋亡的影响。方法:设计、合成有效的mcl-1特异性siRNA序列,体外转染HepG2细胞;通过绘制细胞生长曲线和MTT实验检测mcl-1特异性siRNA对HepG2细胞增殖的影响;通过Annexin V/PI双标记流式细胞仪检测mcl-1特异性siRNA对HepG2细胞凋亡率的影响。结果:通过绘制细胞生长曲线发现,mcl-1特异性siRNA能够抑制HepG2细胞的增殖(P<0.05),MTT实验提示转染mcl-1特异性siRNA 24 h、48 h、72 h后,HepG2细胞存活率均显著下降(P<0.05);流式细胞仪检测分析发现,转染mcl-1特异性siRNA后Annexin V+/PI-细胞百分率显著增高(P<0.01),提示mcl-1具有促进HepG2细胞凋亡的作用。结论:Mcl-1蛋白具有促进肝癌细胞增殖,抑制肝癌细胞凋亡的作用,这种分子特性符合肿瘤靶向治疗的要求,Mcl-1可能成为肝癌靶向治疗的潜在靶点。

Objective： Targeted therapy is the central issue ofhepatocellular carcinoma cells （HCC） currently. The roles of HCC related gene mcl-1 in HCC proliferation and apoptosis are unclear. In this study, the effects of mcl-1 specific siRNA on proliferation and apoptosis of HepG2 cells in vitro will be discussed. Methods： Design and synthesis effective mcl-1 specific siRNA, and mcl-1 specific siRNA was transfect into HepG2 cells in vitro; the effects of mcl-! specific siRNA on proliferation of HepG2 cells were quantified by drawing cell growth curve and MTT assay; the effects of mct-1 specific siRNA on apoptosis of HepG2 cells were observed by Annexin V/PI double staining flow cytometry. Results： Mcl-1 specific siRNA can inhibit proliferation of HepG2 cells （P〈0.05） according to the results of cell growth curve. MTT assay showed that the viability of HepG2 cells was significantly decreased 24 h, 48 h and 72 h after transfected with mcl-1 specific siRNA （P〈0.05）. Annexin V/PI double staining flow cytometry showed that the percentage of Annexin V＋/PI- cells was significantly increased after transfected with mcl-1 specific siRNA （P〈0.01）, which indicated that mcl-1 can promote apoptosis of HepG2 cells in vitro. Conclusion： Mcl-1 can promote proliferation, but inhibit apoptosis of HCC cells in vitro. The molecular characteristics meet the requirements of targeted therapy of tumors. Mcl-1 could be a potential target for HCC targeted therapy.