四嗪二甲酰胺抑制肝癌细胞株HepG2增殖并诱导细胞凋亡的研究

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。

The effects of N,N′-di-(m-methylphenyl)-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide on proliferation and apoptosis in HepG2 cells in vitro

Aim To study the effect of N,N′-di-(m-methylphenyl)-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide(ZGDHu-1) on proliferation inhibition and apoptosis in HepG2 cells in vitro.Method Different concentration of ZGDHu-1 and the different time of cultivate were used to treat HepG2 cell.The proliferation inhibition was analysed by alive cell count,MTT assay and Brdu-ELISA.Cell apoptosis was analysed by cell morphology,DNA agarose gel electrophoresis,DNA content,Annexin-V/PI,cellular DNA feagmentation ELISA and Hoechst33258 labeling method.Results ZGDHu-1 can inhibit HepG2 cell proliferation viability within a certain range of treating time and does,and a majority of HepG2 cells were arrested in G_2/M phase.The HepG2 cells apoposis was comfirmed by type cell morphology and DNA fragment and sub-G1 phase and Hoechst33258 and Annexin/PI Labelingmethod and cellular DNA feagmentation ELISA with a time and does related manner.Conclusion ZGDHu-1 can inhibit proliferation and induced apoptosis of HepG2 cells.