MIIP对膀胱癌T24细胞增殖和迁移侵袭能力的抑制作用及其机制  

The mechanism of MIIP inhibiting proliferation and migration invasion in bladder cancer T24 cells

作　　者：严奉奇 王秦豪[2] 茹懿[2] 马柳疆 陶光晶 张梅[2] 张耀[2] 药立波[2] 李霞[2] 武国军[1] 

机构地区：[1]第四军医大学附属西京医院泌尿外科,陕西西安710032 [2]第四军医大学基础部生物化学与分子生物学教研室,陕西西安710032

出　　处：《现代肿瘤医学》2016年第7期1021-1025,共5页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81272812;81572504)

摘　　要：目的:探讨迁移侵袭抑制蛋白(migration invasion inhibitory protein,MIIP)对膀胱癌T24细胞增殖和迁移侵袭能力的影响及作用机制。方法:设计并合成MIIP过表达质粒,并利用脂质体转染方法上调T24细胞系中MIIP的表达。Western blot及实时定量聚合酶链反应(Real-time PCR)检测过表达MIIP的效果,四甲基偶氮唑蓝法(MTT)及平板克隆法观察MIIP过表达对T24细胞增殖能力的影响,划痕及Transwell实验检验MIIP过表达对T24细胞迁移侵袭能力的影响。Western blot及Real-time PCR检测过表达MIIP后其上皮间充质转化(epithelial-mesenchymal transition,EMT)过程相关标志物钙黏蛋白E(E-cadherin)、钙连蛋白N(Ncadherin)、转录因子Snail蛋白和mRNA变化情况。结果:过表达质粒能有效上调T24细胞系中MIIP蛋白及mRNA的表达,上调MIIP表达后T24细胞增殖能力明显下降,克隆形成能力下降,迁移侵袭能力减弱;上调MIIP表达后E-cadherin表达增加,N-cadherin及转录因子Snail表达减少。结论:过表达MIIP可能通过降解转录因子Snail,从而抑制EMT进程降低膀胱癌T24细胞的增殖及迁移侵袭能力。Objective： To investigate the mechanism of migration invasion inhibitory protein（ MIIP） inhibiting proliferation and migration invasion abilities of T24 cells. Methods： To design and synthesize the MIIP over- expression plasmid P- CMV4- MIIP. Plasmids were transfected into T24 cells and the effects were detected by Western blot and Real- time PCR. The proliferation abilities were tested by MTT assay and colony formation assay. The wound scratch assay and Transwell chamber assay were used to test the migration and invasion abilities. The expression levels of epithelial- mesenchymal transition（ EMT） related molecules E- cadherin,N- cadherin and Snail were detected by Western blot and Real- time PCR. Results： P- CMV4- MIIP could effectively up- regulate the expression of MIIP in T24 cells. Up- regulation of MIIP expression can suppress the cell proliferation and migration invasion capacity of the T24 cells. In addition,the expression of E- cadherin was up- regulated while N- cadherin and Snail were down- regulated when MIIP was increased. Conclusion： MIIP overexpression can inhibit the proliferation and migration invasion capacities by degradation Snail and hindering EMT process in T24 cells.

关 键 词：膀胱癌 迁移侵袭抑制蛋白 上皮间充质转化 迁移侵袭 

分 类 号：R730.21[医药卫生—肿瘤]