Aim To study the effects of new type metal copper complex ( N-Cu ) on proliferation and apoptosis of human hepatoma cell line SMMC-7721 in vitro, and to elucidate the possible mechanism of actions. Methods SMMC-7721 cells were treated with different concentrations of N-Cu ( 0. 3 ~ 24 μmol · L-1 ). The inhibitory effect was examined by MTT assay. The cell cycle and apoptotic rates were detected by flow cytome-try ( FCM ). The expression levels of Bcl-2, Bax and Caspase-3 mRNA and protein were detected by RT-PCR and Western blot. Results N-Cu could remarkably inhibit the growth of SMMC-7721 cells, the suppression was in time-and dose-response relationships. Cell cycle analysis revealed a decreased proportion ofcells in G2/M and S phase, and up-regulation of the rate of G0/G1 , and the apoptotic rate was increased. The expression levels of Bax and Caspase-3 mRNA and protein were up-regulated, but the expression of Bcl-2 mRNA and protein was inhibited in the cells, and all the effects of N-Cu were in a dose-dependent manner. Conclusions N-Cu inhibits cell growth and induces apoptosis in SMMC-7721 cells, arresting cell cycle in G0/G1 phase. The up-regulation of Bax and Caspase-3 and down-regulation of the rate of Bcl-2/Bax may be the most important mechanism of antitumor.%目的 研究新型金属铜络合物(N-Cu)在体外对人肝癌SMMC-7721细胞增殖与凋亡的影响及其作用机制.方法将不同浓度的N-Cu (0.3～24 μmol·L-1)作用于体外培养的SMMC-7721细胞,应用MTT法检测细胞生长抑制率,FCM法检测细胞周期及凋亡率,RT-PCR和Western blot法检测细胞中Bcl-2、Bax、Caspase-3 mRNA 和蛋白表达的变化.结果 N-Cu可明显抑制SMMC-7721细胞的增殖,呈明显的量效与时效关系.随着药物浓度的增加,G0/G1 期的细胞比率上升,G2/M和S期细胞比率下降,并促进凋亡率增加.N-Cu可上调细胞中Bax、Caspase-3基因及蛋白的表达,抑制Bcl-2基因及蛋白的表达,且均呈剂量依赖性.结论 一定浓度的N-Cu可抑制SMMC-7721细胞的增殖并诱导其凋亡,阻滞细胞周期于G0/G1期.上调Bax、Caspase-3基因及蛋白的表达,降低Bcl-2/Bax比值,可能是其诱导细胞凋亡的重要机制.
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