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目的观察四嗪二甲酰胺(ZGDHu-1)体外抑制肝癌细胞株HepG2增殖并诱导细胞凋亡作用。方法将不同浓度的ZGDHu-1与HepG2细胞在体外培养,用台盼蓝染色、MTT法、5′-溴-2′脱氧尿苷(B rdu)-ELISA法观察ZGDHu-1对HepG2细胞增殖的抑制作用;用细胞形态学、DNA凝胶电泳、DNA含量及细胞周期分析、Annexin-V/PI双标记、Ho-echst33258荧光染色和ELISA法测定DNA片段等技术检测细胞凋亡。结果ZGDHu-1能抑制HepG2细胞增殖和活力,呈现作用时间和剂量的量效关系。HepG2细胞与ZG-DHu-1作用后,大部分细胞阻滞于G2-M期;出现典型的细胞形态改变,DNA片断化,亚G1峰检出并增加,Annexin V+/PI-表达升高,细胞内DNA片段含量增加,Hoechst33258荧光染色后出现凋亡细胞的特征性改变等均证实ZGDHu-1能诱导HepG2细胞凋亡。结论ZGDHu-1能抑制HepG2细胞增殖,并可诱导其细胞凋亡。
Objective To investigate the inhibitory effect of ZGDHu-1 on the proliferation and apoptosis of HepG2 hepatoma cell line HepG2 in vitro. Methods Different concentrations of ZGDHu-1 and HepG2 cells were cultured in vitro. The effects of ZGDHu-1 on proliferation of HepG2 cells were observed by trypan blue staining, MTT assay and BrdU-ELISA. Inhibition was detected by cell morphology, DNA gel electrophoresis, DNA content and cell cycle analysis, Annexin-V / PI double staining, Ho-echst33258 fluorescence staining and ELISA assay. Results ZGDHu-1 could inhibit the proliferation and viability of HepG2 cells, showing dose-response relationship between time and dose. After HepG2 cells were treated with ZG-DHu-1, the majority of cells were arrested in G2-M phase. Typical cell morphological changes, DNA fragmentation, sub-G1 peak were detected and increased, Annexin V + / PI- The intracellular DNA content increased, Hoechst33258 fluorescence staining of apoptotic cells and other characteristics of the changes were confirmed ZGDHu-1 can induce HepG2 cell apoptosis. Conclusion ZGDHu-1 can inhibit the proliferation of HepG2 cells and induce its apoptosis.