过表达CDK11^(p58)促进人胰腺导管腺癌MIAPaCa-2细胞增殖  

Overexpression of CDK11^(p58) Promotes the Proliferation of Human Pancreatic Ductal Adenocarcinoma Cell MIAPaCa-2

作　　者：余诗灏[1] 王辉[1] 丁婧[1] 孟雁[1] 

机构地区：[1]中国医学科学院基础医学研究所北京协和医学院基础学院生理学系,北京100005

出　　处：《中国生物化学与分子生物学报》2010年第4期341-346,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助项目(No.30870989);国家重点基础研究发展规划(973计划;No.2006CB503907);科技部专项基金资助项目(No.2004CCA01400);北京市自然科学基金资助项目(No.5062034)~~

摘　　要：CDK11p58属于CDK11/PITSLRE蛋白激酶家族成员,由Cdc2L2编码,是一种重要的细胞周期调控蛋白.为了研究CDK11p58与胰腺癌细胞增殖的关系,我们通过采用脂质体转染真核表达载体及G418筛选的方式,获得了稳定过表达CDK11p58的MIAPaCa-2(人胰腺导管腺癌细胞)单克隆细胞,并通过流式细胞分析、MTT检测及real-time PCR的方法检测了细胞周期、细胞增殖能力及G1/S期相关调控基因的转录水平.结果显示,该单克隆细胞(实验组)与空载体组细胞和空白对照组细胞相比G1期细胞比例明显下降(P<0.01),S期细胞比例明显上升(P<0.01);细胞增殖能力明显提高(P<0.01);cyclin D1、cyclin D3、p21基因mRNA水平较两组对照细胞明显升高(P<0.01).提示过表达的CDK11p58通过上调cyclin D1、cyclin D3和p21基因的mRNA水平促进MIAPaCa-2细胞通过细胞增殖的关键限速点G1/S期,加快细胞增殖.CDK11^p58,a CDK11 /PITSLRE protein kinase family member,is encoded by Cdc2L2,and it plays a crucial role in cell cycle regulation.In order to investigate the relation between CDK11p58 and the proliferation of human pancreatic carcinoma cells,the recombinant eukaryotic expressional plasmid pcDNA3.0-HA-p58 was introduced into human pancreatic ductal adenocarcinoma cell line MIAPaCa-2 by Lipofectamine2000 and the cell was selected by G418.The cell clones which stably overexpress CDK11^p58 were identified by Western blot.Cell cycle,cell proliferation and G1 /S related gene mRNA levels were analyzed by flow cytometry assay,MTT assay and real-time PCR assay,respectively.Results showed that the S phase cell proportion,the proliferation activity and the mRNA level of cyclin D1,cyclin D3,p21 gene in pcDNA3.0-HA-p58 transfected cell were significantly increased compare with the cells in vector group and control group（P 〈0.01）.These results indicated that overexpression of CDK11^p58 could accelerate MIAPaCa-2 cell G1 /S progression and promote its proliferation.

关 键 词：CDK11p58 Cdc2L2 胰腺癌 MIAPACA-2 

分 类 号：Q71[生物学—分子生物学]