SHP-2酪氨酸磷酸酶对肝癌细胞增殖的影响  

Effects of SHP-2 tyrosine phosphatase on proliferation of liver cancer cells

作　　者：杨翠[1] 汪心怡[1] 李菲菲[1] 瞿成奎 汪思应[1] 

机构地区：[1]安徽医科大学,合肥230032 [2]Case Western Reserve University

出　　处：《山东医药》2013年第36期4-6,10,I0002,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30873046;30973424)

摘　　要：目的建立SHP-2野生过表达型(WT)及激活突变型(MT)稳定转染的肝癌HepG2细胞系,研究SHP-2激活突变及野生过表达对肝癌细胞增殖能力的影响。方法将已经构建成功的WT SHP-2(WT组)和MT SHP-2(MT组)及空载体pcDNA3.1(空载组)分别转染到人肝癌HepG2细胞中,同时设未转染细胞作对照组;Western blot法检测细胞中SHP-2蛋白的表达水平;MTT法检测细胞增殖情况;平板克隆和软琼脂集落形成实验检测细胞集落、克隆形成能力。结果成功的将SHP-2突变型和野生型真核表达载体转染到人肝癌HepG2细胞中;MTT结果显示,对照组和空载组细胞增殖速度接近且增殖慢,而MT组和WT组细胞增殖速度明显加快,WT、MT组与空载组、对照组比较,P均<0.01;细胞克隆形成的数量多于空载组和对照组,P均<0.01。结论成功构建了WT及MT稳定表达SHP-2的HepG2细胞株;SHP-2激活突变及野生过表达促进肝癌细胞增殖能力。Objective To construct the HepG2 cell line stably transfected by SHP-2 over-expression wild-type （WT） and gain-of-function mutation （ MT）, and to investigate the effects of SHP-2 gain-of-Function mutation （MT） and over-ex- pression wild-type （WT） on proliferation capacity of liver cancer cells. Methods The successfully constructed eukaryotic expression vector of SHP-2I）61c/~ （ MT ）, wild-type （WT） and vector pcDNA3.1 were transfected into HepG2 ceils, mean- while, the untransfected ones were used as controls. Western blotting was applied to detect SHP-2 protein expression levels of transfected and untransfected cells. MTI＂ assay was used to detect the cell proliferation. Plate colony and soft-agar colony formation assay was applied to detect cell colony formation ability. Results SHP-2 mutant and wild-type eukaryotic ex- pression vector was successfully transfected into HepG2 cells. MTY assay showed that the proliferation rate of the controi group was similar to that of the no-load group and with slow proliferation, while in the mutant group and over-expression wild-type group, the cell proliferation was significantly faster. Significant differences were found between the latter two groups and the first two groups （P 〈 0.01 ）. Plate colony and soft-agar colony formation experimental results show that the number of colony-forming cells in the mutation group and the over-expression wild-type group was more than that in the no- load group and the control group, and with statistically significant difference （P 〈 0.01 ）. Conclusions HepG2 cell line stably transfected by SHP-2 over-expression wild-type （WT） and gain-of-function mutation （MT） is successfully estab- lished. SHP-2 gain-of-function mutation and over-expression wild-type promote the proliferation capacity of liver cancer cells.

关 键 词：肝肿瘤 SHP-2酪氨酸磷酸酶 激活突变 肝癌细胞 

分 类 号：R818.02[医药卫生—放射医学]