蛋白磷酸酶2A的Cβ亚基在胚胎成纤维细胞凋亡过程中的作用  

Effects of Catalytic Subunit β of Protein Phosphatase 2A on Apoptosis of Embryonic Fibroblasts

作　　者：王庆华[1] 王生存[1] 李斌[2] 刘春[1] 吴刘成[1] 王旭[1] 彭晓清[1] 邵义祥[1] 

机构地区：[1]南通大学实验动物中心,江苏南通226001 [2]南通大学附属医院儿科,江苏南通226001

出　　处：《动物医学进展》2016年第2期64-69,共6页Progress In Veterinary Medicine

基　　金：江苏省高校自然科学研究面上项目(11KJB180010;14KJB180018;15KJB180015)

摘　　要：为探究蛋白磷酸酶2A的Cβ亚基在胚胎成纤维细胞(MEFs)凋亡过程中的作用,用Cβ+/-的杂合子雄鼠与雌鼠1∶2配对,取胚胎期12.5d(E12.5)的胚胎制备MEFs,用PCR、RT-PCR和Western blot方法进行基因型鉴定。同时对P3代MEFs利用流式分选技术检测其细胞凋亡和细胞周期的变化,并采用Western blot检测抗凋亡蛋白Bcl-2的表达。结果采用胰酶消化方法成功获得了MEFs,DNA、RNA和蛋白水平鉴定显示获得了3对3的野生型和Cβ基因敲除MEFs。流式分选发现Cβ基因敲除MEFs和野生型之间细胞分裂期前期(M1期)细胞分别为53.95%±2.81和46.94%±4.17,细胞分裂期后期(M2期)细胞分别为21.14%±2.53和29.83%±3.25。但Bcl-2的蛋白表达量和mRNA表达量没有显著差异。结果表明,虽然蛋白磷酸酶2A的Cβ亚基的缺失并不会导致小鼠胚胎死亡,但Cβ亚基的缺失会轻微影响胚胎成纤维细胞(MEFs)的细胞周期。To investigate the effects of catalytic subunit β of protein phosphatase 2A （PP2A） on ceil cycle and cell apoptosis in mouse embryo fibroblasts （MEFs）, the mouse embryos at E12.5 by mating the heter- ozygotes of Cβ subunit conventional knockout male and female mice at the ratio of 1 ： 2 were obtained. MEFs were prepared by trypsin digestion and genotyped by PCR, RT-PCR and Western blot to identify the genetic basis of each embryo. FACs was carried out to determine the cell apoptosis and cell cycle, meanwhile,the anti-apoptosis protein Bcl-2 was checked by Western blot. Genotyping by PCR, RT-PCR and Western blot showed that both knockout and wildtype MEFs were isolated by trypsin digestion at the ratio of 3 ： 3. The MEFs at M1 stage was 46.94%±4.17 and 53.95±2.81, respectively and M2 stage was 29.83%±3.25 and 21.14%±2.53. There is no significant difference in cell apoptosis between wildtype and Cβ knockout MEFs and slight significant difference between wildtype and knockout MEFs in cell cycle.

关 键 词：胚胎成纤维细胞 细胞凋亡 细胞周期 

分 类 号：S852.33[农业科学—基础兽医学]