摘要
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。
Aim To study the effect of N,N＇-di-（ m-methylphenyl） -3,6-dimethyl- 1,4-dihydro- 1,2,4,5-tetrazine-1,4-diearboamide （ 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, MTI？ assay and Brdu-ELISA. Cell apoptosis was analysed by cell morphology, DNA agarose gel eleetrophoresis, DNA content, Annexin-V/PI , cellular DNA feagmentation ELISA and Hoeehst33258 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 G2/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.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2006年第6期754-759,共6页
Chinese Pharmacological Bulletin
基金
浙江省科技厅资助项目(No2003c33019)
关键词
四嗪二甲酰胺
肝癌
细胞株
HepG2
增殖
细胞凋亡
N, N＇-di-（ m-methylphenyl ）-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide
Hepatic cancer
cell line, HepG2
proliferation
apoptosis
