B19病毒XA株VP1独特区蛋白对Hela细胞凋亡的影响  

Effect of VP1-unique Region Protein of Parvovirus B19-XA on the Apoptosis in Hela Cells

作　　者：迟明[1] 迟海峰[2] 许波[1] 晁玉瑾[1] 郑楠[1] 曹艳华[1] 王曼秋[1] 张国成 

机构地区：[1]济南军区总医院儿科,山东济南250031 [2]总参谋部老干部服务管理局卫生处,北京100191 [3]天津武警儿童医院,天津300100

出　　处：《现代生物医学进展》2014年第33期6446-6449,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81070543)

摘　　要：目的:构建B19病毒XA株VPlu基因的真核表达载体和稳定转染细胞系,探讨B19病毒XA株VPl独特区蛋白对Hela细胞凋亡的影响。方法:采用本课题组前期构建的真核表达载体plRES2-EGFP-Vplu及plRES2-EGFP对照质粒,将其稳定转染至Hela细胞,通过流式细胞术检测EGFP阳性细胞的比例以确定稳定转染的细胞系是否成功构建;提取稳定转染的Hela细胞的总蛋白,通过Western blot检测细胞内凋亡相关基因Caspase3的表达;转染plRES2-EGFP-Vplu及plRES2-EGFP的Hela细胞经过Annexin V和PI的染色后,通过流式细胞术检测其凋亡率。结果:本实验成功构建了plRES2-EGFP-VPlu稳定转染的Hela细胞系,plRES2-EGFP-Vplu稳定转染的Hela细胞中Caspase3的表达较转染对照质粒的Hela细胞明显增加,细胞凋亡率亦显著升高,差异均具有统计学意义(P<0.05)。结论:B19病毒XA株VP1独特区蛋白能够显著促进Hela细胞的凋亡。Objective： To clone VP1-unique region of parvovirus B19-XA and construct eukaryotic expression plasmid and stable transfection cell line, and explore the effect of VP1-unique region of parvovirus B19-XA on the apoptosis in Hela cells. Methods：plRES2-EGFP-VPlu constructed previously was used and transfected to Hela to construct a stable transfection cell line. Flow cytometry was used to detect the transfection rate, and western blot was used to detect the expression of caspase3. Hela cells transfected with plRES2-EGFP-Vplu and plRES2-EGFP were stained with Annexin V and PI and flow cytometry was used to detect the apoptotic rate.Results： A cell line stably expressed VP1-unique region of parvovirus B19-XA was successfully constructed. The expression of Caspase3 and apoptotic rate significantly incireased in the cell line stably expressed VP1-unique region of parvovirus B19-XA（P〈0.05）.Conclusion： VP1-unique region of parvovirus B19-XA could obviously promote the apoptosis of Hela cells.

关 键 词：VPlu基因 稳定转染 细胞凋亡 CASPASE3 

分 类 号：Q75[生物学—分子生物学] Q78