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

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.