p27kipl蛋白与胃癌曲妥珠单抗耐药的相关性  

Correlation between cyclin-dependent kinase inhibitor p27kip1 and trastuzumab-resistance in gastric cancer

作　　者：吴梦莞 郭丽红[1] 左强[1] 

机构地区：[1]南方医科大学南方医院肿瘤科,广州510515

出　　处：《中南大学学报（医学版）》2016年第5期471-476,共6页Journal of Central South University ：Medical Science

基　　金：广东省科技计划项目(2012B031800395)~~

摘　　要：目的:探讨细胞周期蛋白依赖性激酶抑制蛋白p27kip1与胃癌曲妥珠单抗耐药的相关性。方法:以Her-2高表达的人胃癌细胞株NCI-N87为研究对象,采用逐步增加剂量法诱导建立曲妥珠单抗耐药细胞NCI-N87/TR;MTT法检测曲妥珠单抗对细胞的半数抑制浓度(50%inhibitory concentration,IC_(50)),计算耐药指数(resistant index,RI),Western印迹法检测细胞cdk2和p27kipl的蛋白表达;通过cdk2抑制剂Purvalanol A刺激NCI-N87/TR,再以Western印迹法检测相关蛋白表达的变化,MTT法检测耐药细胞对曲妥珠单抗敏感性的改变。结果:与NCI-N87比较,NCI-N87/TR细胞cdk2蛋白表达水平显著升高(P<0.001),p27kip1蛋白水平显著下降(P<0.001);通过Purvalanol A恢复NCI-N87/TR的p27kip1蛋白水平后,NCI-N87/TR对曲妥珠单抗的敏感性显著增强(P<0.001)。结论:p27kip1蛋白水平的下调可能是导致胃癌细胞对曲妥珠单抗耐药的机制之一。Objective：To investigate the correlation between cyclin-dependent kinase inhibitor p27 kipl and trastuzumab-resistance in gastric cancer.Methods：We selected HER2-overexpressed human gastric cancer cell line NCI-N87 to establish trastuzumab-resistant NCI-N87/TR cell line by stepwise exposure to different doses of trastuzumab.The 50%inhibitory concentration（IC_（50）） of trastuzumab and resistance index（RI）were calculated or analyzed by MTT assay.The expression levels of cdk2 and p27 kipl were detected by Western blot.After the treatment with cdk2 inhibitor（Purvalanol A）,the expression levels of relevant proteins in NCI-N87/TR cells were detected by Western blot,and the sensitivity to trastuzumab was analyzed by MTT assay.Results：Compared with NCI-N87 cells,the expression of cdk2 was significantly increased in NCI-N87/TR cells（P〈0.001）,while the expression of p27kip1 showed a significant decrease（P〈0.001）.Restoration of the p27kip1 protein expression by cdk2 inhibitor（Purvalanol A）increased the sensitivity of NCI-N87/TR to trastuzumab.Conclusion：Down-regulation of p27kip1 might be a mechanism for triggering trastuzumab resistance to gastric cancer cell line NCI-N87.

关 键 词：曲妥珠单抗 耐药 胃癌 P27KIP1 CDK2 

分 类 号：R735.2[医药卫生—肿瘤]