下调HIF-2α基因对低氧诱导的肝癌细胞体外生物学行为的影响

Effects of downregulation of HIF-2α gene on biological behaviors of hepatoma cells induced by hypoxia in vitro

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。

bObjective： 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.