鸡贫血病毒VP3基因诱导人肺癌细胞SPC-A1凋亡的作用机制  

The mechanisms of apoptosis in human lung carcinoma cells SPC-A1 induced with VP3 from chicken anemia virus

作　　者：连海[1] 金宁一[2] 李霄[2] 陈立刚[2] 管国芳[2] 孙丽丽[2] 李雪梅[2] 郑洪玲[2] 崔英姬[3] 

机构地区：[1]吉林大学农学部畜牧医学院,长春130062 [2]解放军军事医学科学院解放军基因工程重点实验室,长春130062 [3]吉林大学农学部,长春130062

出　　处：《高技术通讯》2006年第10期1063-1066,共4页Chinese High Technology Letters

基　　金：973计划（G1999011902）资助项目

摘　　要：探索了重组质粒pVVP3对人肺癌细胞SPC-A1的作用及机制.将pVVP3经脂质体介导转染人肺癌细胞SPC-A1,通过MTT方法检测了细胞活力;采用吖啶橙/溴化乙锭（AO/EB）染色分析了肿瘤细胞凋亡;采用罗丹明123和DCFA染色测定了线粒体跨膜电位（△Ψm）和活性氧水平变化;通过底物染色反应检测了Caspase-3活性.检测分析结果表明：重组质粒pVVP3转染人肺癌细胞SPC-A1后,细胞活力明显降低; AO/EB染色可见明显的细胞凋亡形态学变化;与空质粒对照相比,线粒体△Ψm下降（P＜0.01）,活性氧水平升高（P＜0.05）,Caspase-3活性增强（P＜0.01）.以上结果提示,重组质粒pVVP3能够通过上调ROS水平,下调线粒体△Ψm,进而激活Caspase-3,最终诱导人肺癌细胞凋亡.The effect of recombinant plasmid pVVP3 on human lung carcinoma cells SPC-A1 and its mechanism were investigated. The recombinant plasmid pVVP3 was introduced into human lung carcinoma cells SPC-A1 by liposome-mediated transfection. The viability of SPC-A1 cells was determined by MTT staining and apoptosis was detected by AO/EB staining. The alteration of mitochondrial transmembrane potential and ROS level of the cells was detected by flow cytometry （FCM） with rhodamine 123 and DCFA staining. The activation of caspase-3 was assayed by its substrate color reaction. The viability of SPC-A1 cells was decreased significantly and recombinant plasmid could lead to obviously morphological apoptotic changes of SPC-A1 cells by introduction of pVVP3 into SPC-A1 cells for 48h. Mitochondrial transmembrane potential decreased（ P 〈 0.01 ）, ROS level of the cells increased （ P 〈 0.05）, Caspase-3 activity （ P 〈 0.01 ） increased, compared with control cells. The above results showed that the significant apoptosis in SPC-A1 cells can be induced by recombinant plasmid pVVP3. Apoptosis may result from the increase of ROS level, the decrease of mitochondrial transmembrane potential and activation of Caspase-3.

关 键 词：鸡贫血病毒VP3 人肺癌细胞 凋亡 

分 类 号：Q279[生物学—细胞生物学]