下调uPAR表达抑制高侵袭性人膀胱癌细胞系T24的迁移与侵袭  

Down-regulating u PAR inhibits migration and invasion of highly invasive human bladder cancer cell line T24

作　　者：韩俊岭 陈昆[1] 郭亮[1] 马曜辉 韩前河[1] 单中杰[1] 宋东奎[2] HAN Jun-ling;CHEN Kun;GUO Liang;MA Yao-hui;HAN Qian-he;SHAN Zhong-jie;SONG Dong-kui(Department of Urology, People s Hospital of Zhengzhou, Zhengzhou 450003;Department of Urology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450003, China)

机构地区：[1]郑州人民医院泌尿外科,河南郑州450003 [2]郑州大学第一附属医院泌尿外科,河南郑州450003

出　　处：《基础医学与临床》2019年第11期1568-1573,共6页Basic and Clinical Medicine

基　　金：河南省2012年科技发展计划(122102310113);郑州市2015年度科技发展计划(20150064)

摘　　要：目的探究尿激酶型纤溶酶原激活物受体(u PAR)的表达对高侵袭性人膀胱癌细胞系T24体外增殖与侵袭的影响,以及哺乳动物靶向雷帕霉素靶蛋白复合物2(m TORC2)可能的作用。方法设计合成针对u PAR、Rictor、PTEN基因的siRNA和阴性对照NC siRNA。将对数增殖期T24细胞随机分为5组,并设为实验组siu PAR、siRictor、siRictor+siu PAR、siu PAR+si PTEN和对照组NC。用瞬时转染法将以上siRNA及相应组合分别转入5组T24细胞;分别用RT-qPCR和Western blot检测mRNA及蛋白的表达;用MTT法和Transwell侵袭实验分别检测u PAR siRNA对T24细胞增殖及侵袭的影响。结果 T24细胞中u PAR mRNA和蛋白的表达量显著上调(P<0. 05)。沉默u PAR能够显著下调T24细胞中磷酸化Akt Serine-473(P-Akt S473)的表达量(P<0. 05),并抑制T24细胞的迁移与侵袭(P<0. 05)。敲除PTEN基因的T24细胞中,沉默u PAR基因促进Akt S473磷酸化(P<0. 05)。结论沉默u PAR能够抑制T24细胞的迁移与侵袭,在T24细胞中u PAR可能是m TORC2的上游调控因子。Objective To study the effect of urokinase-type plasminogen activator receptor (uPAR) gene expression on the migration and invasion of human bladder cancer cell line T24 and the possible mechanism. Methods uPAR siRNA , Rictor siRNA, PTEN siRNA and NC siRNA were designed, synthesized and transfected into bladder cancer T24 cells. Expression of mRNA and protein was detected by real-time quantitive PCR (RT-qPCR) and Western blot. Cell migration assay and Transwell invasion assay were used to testify the migration and invasion of uPAR-silencing T24 cells. Results uPAR mRNA and protein levels of highly invasive bladder cancer T24 cells showed higher than less aggressive RT4 cells( P <0.05). After T24 cells were transfected with uPAR siRNA, phosphorylation of serine-473, an mammalian target of rapamycin complex 2(mTORC2) target, in extracellular regulated protein kinase B (Akt) was significantly downregulated( P <0.05). On the other hand, silencing uPAR also down-regulated migration and invasion of T24 cells( P <0.05). Moreover, Akt-S473 was neither phosphorylated by uPAR nor mTORC2 in PTEN-negative T24 cells. Instead, silencing uPAR gene derepressed Akt-S473 phosphorylation( P <0.05). Conclusions Silencing uPAR down-regulates migration and invasion of bladder cancer cell. The possible mechanism is proposed that uPAR may function as mTORC2 upstream regulator in T24 cells.

关 键 词：膀胱癌细胞系T24 迁移 侵袭 u PAR m TORC2 

分 类 号：R737.14[医药卫生—肿瘤]