蛋白酪氨酸磷酸酶SHP-2对去血清培养诱导的293T细胞凋亡的作用  

Protein tyrosine phosphatase SHP-2 inhibits apoptosis induced by serum withdrawal in 293T cells

作　　者：王秀丽[1] 董红岩[1] 杨金霞[1] 张明爽[1] 张卓然[1] 吴博[1] 徐长庆[1] 张力[1] 

机构地区：[1]哈尔滨医科大学病理生理教研室,黑龙江哈尔滨150081

出　　处：《中国病理生理杂志》2008年第3期548-551,共4页Chinese Journal of Pathophysiology

基　　金：哈尔滨医科大学留学归国人员科研启动基金资助项目;黑龙江省卫生厅课题资助项目(No.2005-36)

摘　　要：目的:探讨蛋白酪氨酸磷酸酶SHP-2对去血清培养诱导人胚肾293T细胞凋亡的作用。方法:将pIRES-GFP空载体、pIRES-GFP-SHP-2(WT)野生型及pIRES-GFP-SHP-2C459S突变体通过脂质体法转染293T细胞,MTT测定去血清培养对293T细胞增殖的抑制情况,去血清培养293T细胞3 d后,电镜观察超微结构、流式细胞仪检测细胞凋亡率、免疫组织化学方法测定caspase-3表达。结果:去血清培养293T细胞3 d,转染pIRES-GFP-SHP-2(WT)野生型组的293T细胞凋亡率明显低于对照组和pIRES-GFP-SHP-2C459S突变体组;而2转染组超微结构均发生早期凋亡,但并无明显差异;caspase-3免疫组化结果显示SHP-2(WT)野生型组的caspase-3表达率明显低于SHP-2C459S突变体组。结论:SHP-2可能参与到去血清培养诱导细胞凋亡的信号转导通路中并通过caspase-3依赖途径,对细胞的生存起到正向调节作用。AIM： To investigate the potential role of Scy homology 2 domain - containing protein tyrosine phos- phatase 2（ SHP- 2） in apoptosis of 293T cells induced by serum deprivation. METHODS： Transfection of plasmids into 293T cells was performed by liposome protocol. The viability of 293T cells was detected by MTr assay. On day 3 after removing serum the morphological changes of 293T cells were observed by using a transmission electron microscope, apoptosis rate was detected by flow cytometry and the expression of caspase - 3 was measured by immunohistochemisty. RESULTS： The apoptosis rate in wild type SHP - 2 （WT） group was obviously lower than that in the SHP - 2^C459S catalytically inactivated group. At the same time, expression of caspase - 3 showed the similar results. CONCLUSION ： SHP - 2 may actively participate in the signal transduction pathway of apoptosis and play a positive role in cell survival. The underlying mechanism of apoptotic inhibition may be caspase - 3 dependent.

关 键 词：Sic同源2结构域蛋白酪氨酸磷酸酶2 293T细胞 细胞凋亡 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R363[医药卫生—病理学]