三氧化二砷对Bel-7402人肝癌细胞株的促凋亡作用

Arsenic Trioxide Promotes the Apoptosis of Human Hepatocellular Carcinoma Cell Line Bel-7402

目的 :研究三氧化二砷 (As2 O3 )在体外是否具有抗肝癌作用 ,并对其作用机制进行初步探讨。方法 :用不同浓度的As2 O3 处理Bel 740 2人肝癌细胞株及L 0 2正常人肝细胞株 ,应用四甲基偶氮唑盐比色法 (MTT法 )观察As2 O3 对细胞生长的影响 ,并用电镜、流式细胞仪、DNA电泳等检测方法研究其作用机制。结果 :MTT法发现As2 O3 0 .5～ 2 .0mg·L-1的浓度均可显著抑制Bel 740 2人肝癌细胞株的生长 ,而对L 0 2正常人肝细胞株的抑制作用较弱。经As2 O3 作用后 ,透射电镜和扫描电镜观察均显示Bel 740 2细胞发生了显著的凋亡形态改变 ;提取细胞DNA行琼脂糖凝胶电泳可见典型的梯形条带 ;流式细胞仪分析可见有亚G1峰出现 ,凋亡率呈时间和剂量双重依赖性特点。结论 :As2 O3 在体外可显著抑制肝癌细胞株生长 。

Objective The purpose of the experiment was to investigate the anticancer activity of As 2O 3 on liver cancer and its mechanism in vitro .Methods The human hepatocellular carcinoma (HCC) cell line Bel?7402 and the human normal liver cell line L?02 were treated with As 2O 3 in various concentrations for various days,then observed and analysed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) assay,transmission electron microscopy (TEM),scanning electron microscopy (SEM),flow cytometry (FCM),agarose gel electrophoresis.Results By means of MTT assays, it is found that 0.5～2.0?mg·L 1 As 2O 3 could significantly inhibit the growth of Bel 7402 cell line, but the inhibitation was not obvious for L 02 cells. After 72?h of treatment with 2.0 ?mg·L -1 As 2O 3,under the inspection of TEM, most of the HCC cells displayed typically morphological features of apoptosis. Surface protrusions and blebs were well demonstrated by SEM. The DNA ladder was viewed on the agarose gel electrophoresis. Sub G 1 peaks were recorded in DNA histogram of FCM.The apoptosis rate showed a time and dose dependent feature.Conclusion As 2O 3 has obvious antitumor activity on HCC in vitro .Its mechanism may mainly be inducing liver cancer cells to undergo apoptosis.