| 迟明 迟海峰 许波 晁玉瑾 郑楠 曹艳华 王曼秋 张国成.B19 病毒XA株VP1独特区蛋白对Hela细胞凋亡的影响[J].现代生物医学进展英文版,2014,14(33):6446-6449. |
| 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 theApoptosis in Hela Cells |
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| DOI： |
| 中文关键词: VPlu基因 稳定转染 细胞凋亡 Caspase3 |
| 英文关键词: Vplu Gene Stable transfection Cell apoptosis Caspase3 |
| 基金项目:国家自然科学基金项目（81070543） |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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. |
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