PPI1对人宫颈癌HeLa细胞紫杉醇敏感性的影响  

Effect of PPI1 on Sensitivity of Human Cervix Carcinoma Cell Line HeLa to Taxol

作　　者：曾麒燕[1] 曾麟杰 黄勇奇[1] 贺菽嘉[1] 张红[1] 

机构地区：[1]广西医科大学生物化学与分子生物学教研室,南宁530021 [2]广西贵港市骨科医院骨一区,贵港537100

出　　处：《华中科技大学学报（医学版）》2010年第4期489-493,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：广西自然科学基金资助项目(No.0542077)

摘　　要：目的探讨PPI1野生型和活化型突变体的表达对宫颈癌细胞株紫杉醇敏感性的影响。方法建立PPI1野生型和持续活化型突变体稳定表达的HeLa细胞株,用MTT实验、克隆形成实验、流式细胞术等方法观察转染PPI1的HeLa细胞对化疗药紫杉醇的敏感性,通过Western blot分析凋亡相关蛋白的表达及MAPK信号转导通路的活化情况。结果经紫杉醇处理后,PPI1持续活化型突变体的表达明显抑制HeLa细胞的增殖和克隆形成能力;该组细胞凋亡率及G2/M期细胞所占比例明显升高;Western blot分析表明,该组细胞促凋亡分子Bax的表达、Caspaes-3和Caspaes-8的活化及Bcl-2和p38的磷酸化程度明显增强。结论 PPI1持续活化型突变体的表达可明显增强HeLa细胞对紫杉醇的敏感性,其中涉及到Bax表达和Bcl-2磷酸化增强,Caspaes-3、Caspaes-8的活化增强和p38信号转导通路的活化增强。Objective To study the effects of PPI1 gene on the sensitivity of human cervix carcinoma cell line HeLa to taxol and the possible mechanism. Methods Plasmids containing wild type or activated mutant of PPI1 gene were transfected into human cervix carcinoma cell line HeLa. G418 was used to select the cell clones constantly expressing PPI1 protein（HeLa-PPI1/ wt, HeLa-PPI1/SI1）. The expression of PPI1/wt fusion protein and PPI1/sl1 was confirmed by RT-PCR or Western blot. The sensitivity of cells to taxol was examined by using MTT assay,colony formation assay,and flow cytometry. Immunoblotting was used to determine apoptosis-related proteins and the effects of PPI1 on MAPK signal transduction by treating the cells with taxol. Results PPI1/wt or PPI1/sI1 was efficiently expressed in HeLa cells. After PPI1/sll transfected HeLa cells were treated with taxol,proliferation and clonogenic ability was impaired markedly,meanwhile the apoptotic cells and G2/M phase cells increased significantly,and moreover, the expression of Bax and phosphorylation of Bcl-2, as well as activation of Caspase-3, Caspase-8 and p38 was enhanced. Conclusion The expression of PPI1/sI1 could enhance the sensitivity of HeLa cells to taxol, which might involve in enhancing the phosphorylation of Bcl 2,activation of Caspase-3,Caspase-8 and p38 signal transduction.

关 键 词：Ⅰ型磷酸酶抑制亚基-1 紫杉醇 细胞凋亡 细胞周期 信号转导 

分 类 号：R349.5[医药卫生—基础医学]