亚砷酸对肝癌BEL-7402细胞增殖、凋亡及其Bcl-2表达的影响

Effects of arsenic acid on proliferation,apoptosis and Bcl-2 expression of BEL-7402 cells

目的探讨亚砷酸(AA)对人肝癌BEL-7402细胞增殖、凋亡及其Bcl-2表达的影响。方法采用MTT比色法检测AA作用后的BEL-7402细胞增殖抑制率,流式细胞术检测BEL-7402细胞周期及凋亡细胞,HE染色法观察凋亡细胞的形态,RT-PCR检测BEL-7402细胞的Bcl-2mRNA,免疫组化法检测细胞的Bcl-2蛋白。结果1.0～8.0μmol/L的AA可使BEL-7402细胞增殖抑制率上升,能诱导BEL-7402细胞凋亡并阻滞细胞周期于S、G2/M期,呈剂量依赖性;8.0μmol/L的AA作用BEL-7402细胞48h后,细胞呈现明显的凋亡形态改变,其Bcl-2mRNA及蛋白表达明显减弱。结论AA体外有抑制BEL-7402细胞增殖及诱导凋亡的作用,且呈时间、剂量依赖性,其作用机制可能与降低其Bcl-2表达有关。

Objective To investigate the effects of arsenious acid （AA） on proliferation, apoptosis and Bcl-2 expres- sion of HCC cell line BEL-7d02. Methods BEL-7402 cells were treated by AA, the proliferation were measured by MTT, the cell cycle and apoptosis were detected by flow cytometry, the morphologic changes of cell apoptosis were observed by HE staining, the Bcl-2 mRNA was determined by RT-PCR, the Bcl-2 protein was measured by immunohistochemical method. Results After treatment with AA in concentrations from 1.0 to 8.0 μmol/L, BEL-7402 cells＇ inhibitive rate increased, the cell apoptosis and arrest the ceils at S, G2/M phase in a dose dependent manner. After treatment with 8.0 μmol/L AA for 48 h, BEL-7402 cells showed evident morphologic changes of apoptosis, and Bcl-2 mRNA and protein expressions were significantly down-regulated. Conclusions In vitro, AA could inhibit growth and induce apoptosis of human hepatocarcinoma cells in a time and dose dependent manner, to lower Bcl-2 expression might be partly the mechanism.