XIAP、OMI/HTRA2蛋白对HeLa细胞的影响及两种蛋白间相互作用

目的探讨XIAP、Omi/HtrA2、米非司酮对HeLa细胞凋亡的影响以及XIAP、Omi/HtrA2的相互作用。方法细胞转染、米非司酮诱导HeLa细胞凋亡,利用共聚焦显微镜及流式细胞术观察XIAP、Omi/HtrA2在细胞内的表达和定位,并检测其对HeLa细胞凋亡影响的情况,使用共聚焦显微镜检测XIAP、Omi/HtrA2的相互作用。结果 (1)在转染了重组质粒pDsRed2-XIAP、pDsRed2-Omi/HtrA2的HeLa细胞中,加入米非司酮后其凋亡率分别是:只加米非司酮组为46.16%;米非司酮+转染质粒pDsRed2-XIAP为26.41%;米非司酮+转染质粒pDsRed2-Omi/HtrA2为78.85%。(2)利用共聚焦显微镜检测XIAP、Omi/HtrA2在HeLa细胞中存在能量共振转移现象。结论米非司酮能促进HeLa细胞凋亡;DsRed2-XIAP融合蛋白能够抑制由米非司酮诱导的HeLa细胞凋亡;DsRed2-Omi/HtrA2融合蛋白能够促进HeLa细胞凋亡,米非司酮能增加Omi/HtrA2促凋亡作用。在活HeLa细胞中,利用荧光共振能量转移技术直接证实XIAP与Omi/HtrA2在部分细胞中存在相互作用。

The effects of fluorescent protein recombinant plasmid with XIAP and OMI/HTRA2 on HeLa cells

Purpose To investigate the effect of XIAP,Omi/HtrA2 and mifepristone on the apoptosis of HeLa cells,and to explore the interaction of XIAP and Omi/HtrA2.Methods Expression,intracellular localization and function of the XIAP,Omi/HtrA2 in HeLa cells were observed by confocal microscopy after transfection.HeLa cells were induced by mifepristone.At the same time,the apoptosis were analyzed for different groups by flow cytometry.The interaction of XIAP and Omi/HtrA2 were analyzed by confocal microscopy.Results (1)HeLa cells induced by mifepristone showed partly apoptosis(46.16%).But,HeLa cells transfected with pDsRed2-XIAP and induced by mifepristone showed 26.41% apoptosis rate.HeLa cells transfected with pDsRed2-Omi/HtrA2 and induced by mifepristone showed 78.85% apoptosis rate.(2)Confocal microscopy detection showed XIAP and Omi/HtrA2 resonance energy transfer phenomena in HeLa cells.Conclusion Mifepristone can promote apoptosis of HeLa cells.The DsRed2-XIAP fusion protein can inhibit apoptosis of HeLa cells by inducing of mifepristone.The DsRed2-Omi/HtrA2 fusion protein can promote apoptosis of HeLa cells by inducing of mifepristone.We demonstrate that the EGFP-Omi/HtrA2 and DsRed2-XIAP fusion protein exist interaction in living HeLa cell.