微小RNA-497对膀胱癌上皮-间充质转化过程的影响  

Effect of microRNA -497 -5p on epithelial -to -mesenchymal transition in bladder cancer

作　　者：杨成林 王尉[1] 聂海波[1] 肖远松[1] 胡小燕[1] 张小明[1] 汪帮琦[1] 周五二 Yang Chenglin;Wang Wei;Nie Haibo;Xiao Yuansong;Hu Xiaoyan;Zhang Xiaoming;Wang Bangqi;Zhou Wuer(Department of Urology,Guangzhou General Hospital of Guangzhou Military Command,Guangzhou 510010,China;Graduate School of Southern Medical University,Guangzhou 510.515,China)

机构地区：[1]广州军区广州总医院泌尿外科,510010 [2]南方医科大学研究生学院,广州510515

出　　处：《中华实验外科杂志》2018年第9期1595-1597,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81372744）

摘　　要：目的观察微小RNA（miRNA，miR）-497在膀胱癌细胞株中的表达，探讨miR-497对膀胱癌上皮一问充质转化（EMT）过程的影响。方法采用实时定量聚合酶链反应（RT-qPCR）检测miR-497在膀胱细胞株和正常膀胱黏膜组织中的表达。将miR-497mimics／inhibitor分别瞬时转染至膀胱癌EJ细胞中，通过显微镜细胞形态学观察、细胞划痕实验、Transwell侵袭实验，分析过表达或下调内源性miR-497的表达对EJ细胞形态、迁移和侵袭的影响；Westernblot法检测EMT标志物相关蛋白表达的变化。结果与膀胱永生化上皮细胞（SV-HUC-1），miR-497在5株膀胱癌细胞株中均呈低表达状态（P=0．000）。过表达miR-497后，EJ细胞表现为上皮样细胞形态，EJ细胞的迁移率为（25．33±6．11）％，较其阴性对照组（74．00±3．61）％降低2．96倍，侵袭细胞个数为（109±3）个，较其阴性对照组[（219±17）个]降低2倍；而抑制miR-497表达后，EJ细胞表现为问质样细胞形态，EJ细胞的迁移率为（74．00±3．61）％，较其阴性对照组[（51．33±6．51）％]增加1．45倍，侵袭细胞个数为（320±14）个，较其阴性对照组[（248±16）个]增加1．29倍。Westernblot结果显示：与阴性对照组比较，过表达miR-497可使EJ细胞的E-钙黏素（E-cadherin）表达增加（P=0．000），N-钙黏素（N-cadherin）和波形蛋白（Vimentin）表达减少（P=0．001，P=0．000）。结论miR-497可能参与膀胱癌EMT过程的调节。Objective To observe the expression of microRNA- 497 （miR- 497 ） in bladder cancer cells and investigate its possible functions in epithelial- mesenchymal transition （EMT） of bladder cancer. Methods The expression of miR -497 in bladder cells and human normal bladder tissue were measured by real- time quantitative polymerase chain reaction （RT- qPCR）. MiR- 497 mimics and miR - 497 inhibitor were transfected into bladder cancer EJ cells and RT - qPCR was used to analyze the effect of transfection. Then, morphological changes of cells were observed under the inverted microscope and the wound healing assay, transwell invasion assay were used to detect the cell migration and invasion of bladder cancer EJ cells transfected by miR- 497 mimics and miR- 497 inhibitor, cytometry. Finally, Western blotting was applied to detect the levels of EMT - related proteins. Results MiR - 497 levels were obviously down - regulated in bladder cancer cells （5637, EJ, BIU - 87, TCCSUP and T24） in com- parison to human bladder pillon biochemical cell （ SV - HUC - 1 ）, the difference was statistically signifi- cant （ P=0.000）. Compared with negative control, overexpression of miR - 497 significantly suppressed EJ cells migration [ （74.00±3.61）% vs. （25.33±6. 11）%] and invasion [ （219±17） cells vs. （109 ± 3） cells] , and EJ cells turned into epithelioid cells morphology, when compared with the control group. Moreover, opposite results were obtained when miR -497 inhibitor was transfected into EJ cells. Compared with mimics NC, the protein expression of E - cadherin was npregulated （ P=0.000）, while N - cadherin and vimentin downregulated （P=0.001, P = 0.000） in miR -497 mimics transfection group. Conclusion MiR -497 may play an important role of EMT in bladder cancer.

关 键 词：微小RNA-497 膀胱癌 迁移 侵袭 上皮-间充质转化 

分 类 号：R737.14[医药卫生—肿瘤]