活细胞中丝裂霉素诱导肺腺癌细胞凋亡机理的分析  

An Analysis of MMC-induced ASTC-a-1 Apoptosis in Living Cells

作　　者：王会营[1] 陈同生[1] 孙磊[1] 

机构地区：[1]华南师范大学激光生命科学研究所暨激光生命科学教育部重点实验室,广东广州510631

出　　处：《激光生物学报》2008年第4期435-439,共5页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(30670507;60378043;30470494);广东省自然科学基金项目(015012)

摘　　要：为了分析丝裂霉素(mitomycin C,MMC)引起的肺腺癌细胞(ASTC-a-1)凋亡,实验通过CCK-8试剂盒检测不同浓度MMC对ASTC-a-1活性的影响,选择10μg/mL的MMC处理ASTC-a-1。利用Hochest 33258染色观察MMC引起的ASTC-a-1凋亡,发现MMC可引起细胞缩小,核浓缩。为了进一步研究MMC引起凋亡的途径,通过脂质体转染pSCAT3质粒,利用光漂白技术和光谱技术观察Caspase-3是否活化;通过转染Bax和Ds-Red质粒观察Bax在凋亡过程中的位置变化及与线粒体的关系。结果显示:MMC可以诱导ASTC-a-1细胞内Caspase-3活化,并使Bax向线粒体转移聚集。在活细胞中证实Caspase-3和Bax参与了MMC引起的ASTC-a-1凋亡。To investigate mitomycin C （MMC）-induced apoptosis of ASTC-a-1, the cells were treated with MMC at differcut couceutration and their activity was assessed by cell counting kit （ CCK-8）. Based on results of CCK-8, cells were treated with 10 μg/ml, of MMC. Hochcst 33258 has been used to verifiy the MMC-induced cell apoptosis and the results indicated that MMC induced cell and nuclear shrinked. For advanced investigation of the apoptosis pathway induced by MMC, human lung adenocarcinoma cells （ ASTC-a-1 ） was transfected with SCAT3 plasmid, a fluorescence resonance energy transfer （FRET） plasmid. Photobleaching and emission spectrum techniques have been used to monitor the activation of Caspase-3 in living cells. ASTC-a-1 was transfected with plasmid Bax and Ds-Red to monitor the translocation of Bax into mitochondria during apoptosis induced by MMC. Our results demonstrate that both Caspase-3 activation and Bax are involved in the MMC-induced apoptosis.

关 键 词：丝裂霉素 肺腺癌细胞 凋亡 CASPASE-3 BAX 

分 类 号：Q25[生物学—细胞生物学]