三氧化二砷诱导人肝癌SMMC-7721细胞凋亡及对OPN表达的影响

Apoptosis induced by arsenic trioxide and its effects on OPN expression in human hepatocellular carcinoma SMMC-7721 cell

目的探讨三氧化二砷(As2O3)治疗肝癌的可行性及机制。方法将一定浓度梯度的As2O3与人肝癌细胞株SMMC-7721孵育后,采用MTT法、荧光显微镜及流式细胞仪检测细胞增殖与凋亡变化,逆转录聚合酶链反应(RT-PCR)检测肝癌细胞株中骨桥蛋白基因(OPN mRNA)表达水平。结果As2O3作用后SMMC-7721细胞生长明显受抑,且呈时间—浓度依赖性;荧光显微镜下细胞呈典型的凋亡形态学改变;在流式细胞仪上可见"凋亡峰",细胞周期阻滞于G2/M期;细胞OPN mRNA表达阳性,OPN mRNA表达水平明显下调。结论As2O3体外能有效抑制肝癌细胞株生长;其机制可能为诱导细胞凋亡、下调OPN mRNA表达。

Objective To investigate the feasibility and mechanism of the arsenic trioxide （As2O3 ） for the treatment of human hepatocellular carcinoma. Methods The human hepatocellular carcinoma cell line SMMC-7721 were cultured with As2O3 in certain concentration gradient, then the proliferation and apoptosis of SMMC-7721 cell were detected by MTT, fluorescent microscope and flow cytometry（FCM）. The gene expression of osteopontin was detected by RT-PCR. Results The proliferation of SMMC-7721 cell were inhibited significantly after the treantment of As2O3 ,the inhibitory rate was depended on the concentration of As2O3 and treatment time. The SMMC-7721 cell nucleus showed the typical change of apoptpsis morphology. The ＂apoptotic peak＂ was seen visibly. SMMC-7721 cell cycle was arrested at G2/M phase. The expression of osteopontin mRNA was positive in SMMC-7721 and As2O3 could down-regulated that. Conclusion As2O3 has obvious antitumor activity on hepatocellular carcinoma cell lines, the mechanism may be induce the apoptosis of tumor cells, and down-regulate the expression levels of osteopontin mRNA.