H_2O_2对人肝癌细胞Hep-G2凋亡的影响  

Effects of Hydrogen Peroxide on Apoptosis of Hep-G2 Cells

作　　者：伍春莲[1] 房守敏[1] 耿文峰[1] 张思楠[1] 孙警辉[1] 李祥会[1] 

机构地区：[1]西华师范大学生命科学学院西南野生动植物资源保护教育部重点实验室,四川南充637009

出　　处：《西华师范大学学报（自然科学版）》2010年第4期342-346,共5页Journal of China West Normal University(Natural Sciences)

基　　金：四川省教育厅青年基金项目(07ZB089);西华师范大学科研启动基金资助项目(06B026)

摘　　要：探讨不同浓度H2O2对人肝癌细胞株Hep-G2细胞凋亡的影响.使用不同浓度(0.01、0.05、0.10、0.20、0.50mmol/L)的H2O2作用于Hep-G2细胞12h,通过丫啶橙-碘化丙啶(AO/PI)双重荧光染色法对Hep-G2细胞的形态变化进行观察;再进行DNA的提取和琼脂糖凝胶电泳观察DNA断裂情况.结果表明:不同浓度的H2O2处理后在荧光显微镜下可以区分正常对照细胞(0mmol/L)、染色质凝聚的早期凋亡细胞(0.01、0.05mmol/L)、细胞质皱缩和细胞核解体的晚期凋亡细胞(0.10、0.20mmol/L)和降解细胞(0.50mmol/L),在一定时间内随H2O2浓度的增加而增大,呈现浓度依赖性.DNA电泳结果显示发生凋亡的细胞基因组DNA形成弥散条带,从而表现出体外细胞凋亡的特征.因此,H2O2可以诱导Hep-G2细胞凋亡.To investigate the effects of different concentrations of hydrogen peroxide on apoptosis in Hep-G2 cells,Hep-G2 cells were treated 12 hours by different concentrations of hydrogen peroxide that were 0.01、0.05、0.10、0.20、0.50mmol/L and the acridine AO/PI double staining fluorescence was used for Hep-G2 cells line to detect the morphological changes of apoptotic cells under fluorescence microscope.Genomic DNA of Hep-G2 cells was extracted for testing base pair fragments.The results showed the cells could be classificated into： normol cells（0mmol/L）;early apoptotic cells with chromatin agglomerating（0.01,0.05mmol/L）;last apoptotic cells with cytoplasm buckling and cell nucleus disassembly（0.10,0.20mmol/L） and degradationed cells under the fluorescence microuscopy,and in a certain period they increased with increasing concentration of H2O2 which showed dose-dependent.The genomic DNA of apoptotic Hep-G2 cells could be degradated into disperse DNA bands,which showed the characteristics of cell apoptosis in vitro.Therefore,human hepatoma cells Hep-G2 cell could be induced to apoptosis by H2O2.

关 键 词：H2O2 Hep-G2细胞 AO/PI 细胞凋亡 DNA 

分 类 号：Q255[生物学—细胞生物学]