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目的探讨藤黄酸(GA)对人肝癌Bel-7402细胞的诱导凋亡和增殖抑制作用。方法采用四甲基偶氮唑蓝(MTT)比色法检测细胞生长的抑制作用,流式细胞术检测细胞凋亡率及细胞周期变化,RT-PCR和Western blot分别检测Bcl-2、Bax mRNA及蛋白质的表达量。结果经1、2、4μmol/L GA作用Bel-7402细胞24、48、72h后,细胞凋亡率有不同程度上升,呈现剂量和时间依赖性细胞增殖抑制作用;细胞凋亡率随着GA浓度的增加而逐步增高,且产生G2/M期细胞周期阻滞和凋亡;Bax mRNA和蛋白表达上调,而Bcl-2mRNA及蛋白表达降低。结论 GA对Bel-7402细胞有显著的诱导凋亡及增殖抑制作用。其作用机制可能与上调Bax基因、下调Bcl-2基因的表达以及对细胞周期的阻滞有关。
Objective To investigate the inhibitory effect of gambogic acid (GA) on the apoptosis and proliferation of human hepatoma Bel-7402 cells. Methods MTT assay was used to detect the cell growth inhibition. Flow cytometry was used to detect the apoptosis rate and cell cycle changes. The expressions of Bcl-2 and Bax mRNA were detected by RT-PCR and Western blot, respectively And the amount of protein expression. Results Bel-7402 cells treated with 1, 2, 4μmol / L GA for 24,48,72h, the apoptotic rate increased to some extent, showing a dose-dependent and time-dependent inhibition of cell proliferation; with the GA concentration And increased G2 / M phase cell cycle arrest and apoptosis; Bax mRNA and protein expression increased, while Bcl-2 mRNA and protein expression decreased. Conclusion GA can induce apoptosis and proliferation in Bel-7402 cells. Its mechanism may be related to up-regulation of Bax gene, down-regulation of Bcl-2 gene expression and cell cycle arrest.