tBid基因的表达及其对骨肉瘤SOSP-9607细胞的促凋亡作用  

Expression of tBid gene and it' s pro-apoptotic effect on osteosarcoma SOSP-9607 cells

作　　者：纪振钢[1] 裘秀春[1] 杨彤涛[1] 龙华[1] 周本根[1] 文艳华[1] 许彦鸣[2] 杨安钢[3] 范清宇[1] 

机构地区：[1]第四军医大学唐都医院骨科,陕西西安710038 [2]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710032 [3]第四军医大学基础部免疫学教研室,陕西西安710032

出　　处：《现代肿瘤医学》2007年第5期598-600,共3页Journal of Modern Oncology

基　　金：国家自然科学基金重点项目(NO.30330610);国家自然科学基金面上项目(No.30471988);中国博士后基金资助项目(No.2005038259)

摘　　要：目的:观察tBid基因在骨肉瘤细胞中的表达及其对转染的SOSP-9607细胞的凋亡作用。方法:采用脂质体转染的方法,将tBid基因转染骨肉瘤细胞,间接免疫荧光法检测目的蛋白的表达和细胞形态学变化,进一步电镜观察其超微结构改变。MTT法检测目的基因转染后细胞的增殖情况,通过Annexin V、染色流式细胞仪检测来观察其促凋亡作用。结果:转染SOSP-9607细胞后,间接免疫荧光染色检测tBid的表达,细胞出现凋亡。电镜观察呈现出典型的凋亡特征。MTT实验发现细胞的增殖被明显抑制。Annexin V染色流式细胞仪检测可见明显的凋亡细胞,与对照组细胞(凋亡率为6%)相比,其凋亡率为35.8%。结论:tBid基因可以在转染的SOSP-9607细胞中表达,并可抑制转染细胞的生长,诱导细胞发生凋亡。Objective： To observe the pro - apoptotic effect of tBid protein on osteosarcoma SOSP -9607 cells. Methods：tBid gene was transfected into osteosarcoma cells under lipofectamine mediation. The effect of target gene expression on morphology and growth status of SOSP -9607 were observed by fluorescence and electron microscopy and was analyzed by Annexin V - FITC staining and MTF assay. Results： After osteosarcoma cells transfected with tBid gene, the target protein was expressed and followed by the cell shrinkage and cell death. The transfected cells presented the typical characteristics of apoptosis as detected by electron microscopy . The proliferation of tBid gene - transfected SOSP -9607 cells was lower than that of mock - transfected cells. Annexin V - FITC staining revealed that the percentages of apoptotic cells in the transfectants of tBid genes were 35.8%, compared to 6% in the control cells. Conclusion：The expression of tBid can effectively induce apoptosis of osteosarcoma cells.

关 键 词：TBID 骨肉瘤SOSP-9607细胞 凋亡 

分 类 号：R730[医药卫生—肿瘤]