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目的:研究HIF-1α对常氧状态下培养的人非霍奇金淋巴瘤(NHL)细胞凋亡的作用及其调控机制。方法:以HIF-1α抑制剂Echinomycin(EC)处理人NHL细胞,采用Annexin-Ⅴ染色方法检测肿瘤凋亡水平的变化,采用蛋白质印迹法检测各细胞系中Survivin及凋亡相关蛋白Bcl-2和caspase-3的表达水平;进一步应用HIF-1α反义质粒转染肿瘤细胞,检测转染后Sur-vivin蛋白表达水平。结果:HIF-1α抑制剂EC能够诱导NHL细胞的凋亡(P<0.05),具有时间和剂量依赖性。抑制HIF-1α的表达明显抑制了肿瘤细胞中Survivin蛋白表达水平,同时,凋亡相关蛋白Bcl-2表达减低而Caspase-3蛋白表达增加,与对照组相比差异均具有统计学意义,P<0.05。转染反义HIF-1αsiRNA后肿瘤细胞中Survivin蛋白表达水平明显下降(P<0.05)。结论:抑制HIF-1α能够诱导NHL细胞凋亡,其机制可能与下调Survivin蛋白表达水平有关。
Objective: To investigate the effect of HIF-1α on the apoptosis of human non-Hodgkin’s lymphoma (NHL) cultured in normoxia and its regulatory mechanism. Methods: The human NHL cells were treated with HIF-1α inhibitor Echinomycin (EC), and the changes of apoptosis were detected by Annexin-Ⅴ staining. The expressions of Survivin, Bcl-2 and Bcl-2 in each cell line were detected by Western blotting The expression of Survivin protein was detected by Western blotting. The expression of Survivin protein was detected by Western blotting. Results: HIF-1α inhibitor EC could induce apoptosis of NHL cells (P <0.05) in a time-and dose-dependent manner. Inhibition of HIF-1α expression significantly inhibited the expression of Survivin protein in tumor cells, meanwhile, the expression of Bcl-2 protein and Caspase-3 protein were increased, the difference was statistically significant (P < 0.05. Survivin protein expression was significantly decreased in tumor cells transfected with antisense HIF-1αsiRNA (P <0.05). Conclusion: Inhibition of HIF-1α can induce apoptosis of NHL cells, which may be related to down-regulation of Survivin protein expression.