MicroRNA-497对人膀胱癌EJ细胞增殖和凋亡的影响  

Effect of micro RNA-497 on the proliferation and apoptosis of baldder cancer cell line EJ

作　　者：杨成林 王尉[2] 周五二 聂海波[2] 罗敏[3] 张长征[2] 曾钦松 

机构地区：[1]南方医科大学研究生学院,广州市510515 [2]南方医科大学附属广州军区广州总医院泌尿外科,广州市510010 [3]南方医科大学附属广州军区广州总医院妇产科,广州市510010

出　　处：《实用医学杂志》2017年第20期3348-3353,共6页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:81372744);广东省自然科学基金资助项目(编号:2016A030313616);广州市科技基金资助项目(编号:201607010184)

摘　　要：目的探讨microRNA-497(miR-497)对人膀胱癌EJ细胞增殖和凋亡的影响。方法将人膀胱癌细胞株EJ分为4组:miR-497 mimics、mimics NC、miR-497 inhibitor、inhibitor NC;通过脂质体Lipfectami-neTM2000介导将miR-497 mimics、mimics NC、miR-497 inhibitor、inhibitor NC瞬时转染至上述4组细胞中,转染6 h后,在荧光显微镜下观察转染效率;转染48h后,采用qRT-PCR技术检测各组细胞中miR-497的表达;平板克隆实验和噻唑蓝(MTT)实验检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Western blot法检测凋亡相关蛋白Bcl-2和Caspase-3的表达变化。结果荧光显微镜下观察结果显示:各组细胞转染效率均>90%;qRT-PCR结果显示:与各自阴性对照组相比,转染miR-497 mimics组,EJ细胞中miR-497表达明显增加(P<0.01),而转染miR-497 inhibitor后表达明显下调(P<0.01)。过表达miR-497后,EJ细胞的增殖能力较阴性对照组明显减弱(P<0.01),凋亡率明显上升(P<0.01);而抑制miR-497表达后,EJ细胞的增殖能力较阴性对照组明显增强(P<0.05),凋亡率明显降低(P<0.01)。Western blot结果示:与阴性对照组相比,过表达miR-497可使EJ细胞的Bcl-2表达降低(P<0.01),Caspase-3表达增加(P<0.01)。结论miR-497可抑制膀胱癌EJ细胞增殖,促进其凋亡,可能与影响Bcl-2和Caspase-3蛋白表达有关。Objective To investigate the effect of microRNA-497（miR-497）on cell proliferation andapoptosis capability of baldder cancer cell line EJ. Methods EJ human baldder cancer cells were divided into miR-497 mimics group,mimics NC group,miR-497 inhibitor group and inhibitor NC group. Mi R-497 mimics,mimicsNC,miR-497 inhibitor and inhibitor NC were transfected into EJ cells by Lipfectamine^（TM） 2000. The transfection effi-ciency was observed under a fluorescence microscope 6 hours after. And qRT-PCR was used to detected the expres-sion of miR-497 48 hours after. Plate clone formation assay,MTT assay and flow-cytometric analysis of apoptosiswere used to detect the cell proliferation and apoptosis of bladder cancer EJ cells. Western blot was employed to de-termine the protein expressions of bcl-2 and caspase-3. Results Under fluorescence microscope,the efficiencyrate of four groups were over 90%. And qRT-PCR results showed miR-497 expression in EJ cells increased signifi-cantly in miR-497 mimics group than those in the control group（P 0.01）,while miR-497 inhibitor post expres-sion decreased（P 0.01）. Overexpression of miR-497 significantly suppressed EJ cells proliferation（P 0.01）,and prompted EJ cells apoptosis（P 0.01）compared with the control group. Moreover,opposite results were ob-tained when miR-497 inhibitor was transfected into EJ cells.Compared with negative control,the protein expres-sion of Bcl-2 down-regulated（P 0.01）and Caspase-3 up-regulated（P 0.01）in miR-497 mimics transfectiongroup. Conclusions Mi R-497 could suppress the proliferation and promote apoptosis of EJ cells,which might berelated to the protein expression of Bcl-2 and Caspase-3.

关 键 词：膀胱肿瘤 microRNA-497 增殖 凋亡 

分 类 号：R737.14[医药卫生—肿瘤]