【摘要】 目的:探讨下调缺氧诱导因子2α(Hypoxia-inducible factor 2α,HIF-2α)基因对人肝癌细胞株Hep G2低氧状态下增殖、凋亡、细胞周期分布和迁移侵袭力的影响。方法:选取氯化钴(Co Cl2)诱导的人肝癌细胞株Hep G2作为研究对象,构建HIF-2αsiRNA特异性载体,转染低氧环境下的Hep G2细胞,Real-time PCR和Western blot方法分别检测转染前后细胞中HIF-2αmRNA和蛋白表达,MTT方法检测转染前后Hep G2细胞增殖,流式细胞术检测转染前后Hep G2细胞凋亡和细胞周期分布,Transwell试验检测转染前后Hep G2细胞侵袭迁移能力。结果:低氧诱导下,肝癌Hep G2细胞HIF-2α表达显著增多;特异性转染HIF-2αsiRNA于低氧环境下的Hep G2细胞后,HIF-2αmRNA和蛋白表达水平均受到明显抑制、细胞增殖能力降低,凋亡率升高,分布于G0/G1期细胞比率升高,合成期(S)及合成后期(G2/M)细胞比率降低,细胞体外侵袭迁移能力受到明显抑制(P<0.05)。结论:低氧环境下肝癌Hep G2细胞表达HIF-2α增多,特异性siRNA可通过下调低氧环境下Hep G2细胞中HIF-2α基因表达,抑制Hep G2细胞增殖、侵袭迁移,改变细胞周期分布并诱导其凋亡。更多还原

【Abstract】 Objective: To explore the effects of hypoxia-inducible factor 2α genes on under hypoxia on proliferation,apoptosis,cell cycle distribution and migration of invasiveness of human hepatocellular carcinoma cell Hep G2. Methods: Human hepatoma cell line Hep G2 induced by cobalt chloride( Co Cl2) was selected as the research object,construction of siRNA specific carrier HIF-2α,transfection of Hep G2 cells under hypoxia. Real-time PCR,Western blot method in the detection of before and after transfection in each group of HIF-2α mRNA and protein expression; MTT method to detect the proliferation of Hep G2 cells before and after transfection; apoptosis rate and distribution of cell cycle of Hep G2 cells before and after transfection were detected by flow cytometry; Transwell test was used to detect the invasion and migration ability of Hep G2 cells before and after transfection. Results: Under hypoxia,significant increased HIF-2α expression in hepatocellular carcinoma Hep G2 cells. Specific transfection of HIF-2α siRNA in Hep G2 cells after HIF-2α mRNA and protein expression levels were significantly inhibit cell proliferation decreased,apoptosis rate increased in the ratio of G0/G1 phase cells increased synthesis phase( S) and late( G2/M) synthesis cell ratio decreased,which in vitro invasion and migration of cells was inhibited. Conclusion: Expression of HIF-2α increases in hepatocellular carcinoma Hep G2 cells under hypoxia. Specific siRNA can be cut by HIF-2α gene expression in Hep G2 cells under hypoxia,to inhibit Hep G2 cell proliferation,invasion,migration,and change the distribution of cell cycle and induce its apoptosis.更多还原