| 周新文,许雅香.lyGDI在HeLa细胞中的表达与辐射增敏研究[J].中华放射医学与防护杂志,2006,26(6):554-556 |
| lyGDI在HeLa细胞中的表达与辐射增敏研究 |
| lyGDI expression in HeLa cells increased its sensitivity to radiation-induced apoptosis |
| 投稿时间：2006-01-10 |
| DOI： |
| 中文关键词: lyGDI HeLa细胞 辐射增敏 |
| 英文关键词:LyGDI HeLa cell Radiosensitization |
| 基金项目:国家自然科学基金资助项目(30570548);苏州大学医学发展基金资助项目(EE126607) |
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| 中文摘要: |
| 目的 将lyGDI和突变体D19lyGDI转入到不含内源性lyGDI的HeLa细胞中,探讨其是否有凋亡信号转导功能,并分析lyGDI的导入表达能否增加辐射诱导HeLa凋亡的敏感性。方法 用脂质体转染的方法将构建于pCDNA3.1HisA载体上全长的LyGDI和D19lyGDI转入HeLa细胞,用Westernblot检测LyGDI的表达和Xpress标记物,同时,用细胞流式仪和AnnexinV-FITC双染色检测HeLa细胞瞬时转染诱导的细胞凋亡。用G418筛选稳定表达LyGDI的克隆,经12Gy的60Coγ射线辐射处理,Westernblot分析Caspase-3的活性及克隆形成试验来观察LyGDI的导入是否增加HeLa细胞的放射敏感性。结论 IyGDI在HeLa细胞中的瞬时表达诱导了细胞的凋亡,LyGDI具有凋亡信号转导功能。其次,Westernblot检测Caspase-3和克隆形成试验的结果表明LyGDI的导入增加了HeLa细胞辐射凋亡的敏感性。 |
| 英文摘要: |
| Objective In order to confirm whether LyGDI has apoptotic signal transduction function and can increase the apoptotic rate of radiation-induced cell death, the lyGDI and mutant D19lyGDI gene, which constructed with the pCDNA3.1 His A, were transfected into no-endogenous lyGDI HeLa cells. Methods Transient expressions of lyGDI and D19lyGDI in HeLa cells were analyzed by Western blot using anti-mono antibody of LyGDI and Xpress tag. Cell apoptosis was assayed with Annexin V-FITC apoptosis kit. To select stable clone, the transferred HeLa cells had been maintained in G418 medium for 3 weeks, then a cell line, which stably expressed LyGDI and mutant D19lyGDI, was selected. The selected cell line was irradiated with 12 Gy 60Co γ-rays. Caspase-3 activity of the cells was determined by Western blot and cell viability by clone-forming assay after 48 hours post-irradiation culture. Results Western blot and Annexin V-FITC apoptotic analysis revealed that lyGDI and D19lyGDI transient expressions in HeLa cells induced apoptosis; Caspase-3 activity measurement and clone-forming assay showed that lyGDI increased sensitivity to radiation-induced cell apoptosis. Conclusions lyGDI performs function in apoptosis signal transduction, its expression in HeLa cells can increase the sensitivity to radiation-induced cell apoptosis. |
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