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

作　　者：周新文[1] 许雅香[2] 

机构地区：[1]苏州大学放射医学与公共卫生学院,215123 [2]苏州大学生命科学学院

出　　处：《中华放射医学与防护杂志》2006年第6期554-556,共3页Chinese Journal of Radiological Medicine and Protection

基　　金：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)

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。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 D191yGDI gene, which constructed with the pCDNA3.1 His A, were transfected into no-endogenous lyGDI HeLa cells. Methods Transient expressions of lyGDI and D191yGDI 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 D191yGDI, 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 D191yGDI 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.

关 键 词：LYGDI HELA细胞 辐射增敏 

分 类 号：R363[医药卫生—病理学]