MicroRNA-497对人膀胱癌EJ细胞增殖和凋亡的影响

发布时间：2018-03-14 21:03

本文选题：膀胱肿瘤　切入点：microRNA-　出处：《实用医学杂志》2017年20期 　论文类型：期刊论文

【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(P0.01),而转染miR-497 inhibitor后表达明显下调(P0.01)。过表达miR-497后,EJ细胞的增殖能力较阴性对照组明显减弱(P0.01),凋亡率明显上升(P0.01);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(P0.05),凋亡率明显降低(P0.01)。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(P0.01),Caspase-3表达增加(P0.01)。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。
[Abstract]:Objective to investigate the effect of microRNA-497miR-497) on the proliferation and apoptosis of human bladder cancer EJ cells. Methods Human bladder cancer cell line EJ was divided into 4 groups: 1: miR-497 mimics nCmmiR-497 inhibitor inhibitor NC.The miR-497 mimicsmimics NCmiR-497 inhibitory inhibitor NC was transiently transfected into the above 4 cells by liposome Lipfectami-neTM2000 for 6 h after transfection. The transfection efficiency was observed under fluorescence microscope, the expression of miR-497 in each group was detected by qRT-PCR technique after 48 h transfection, the proliferation of cells was detected by plate cloning assay and thiazolyl blue qRT-PCR assay. The expression of apoptosis-related protein Bcl-2 and Caspase-3 was detected by flow cytometry and Western blot. Results the transfection efficiency of each group was 90g / qRT-PCR, which was higher than that of the control group (P < 0.05). After transfection of miR-497 mimics, the expression of miR-497 in EJ cells increased significantly, while that of miR-497 inhibitor decreased significantly after transfection of miR-497 inhibitor. The proliferative ability of EJ cells after overexpression of miR-497 was significantly lower than that of negative control group, and the apoptotic rate of EJ cells increased significantly, while that of EJ cells after inhibiting miR-497 expression was significantly higher than that of negative control group. Compared with the negative control group, the proliferation ability of the cells was significantly increased, and the apoptosis rate was significantly decreased. The results of Western blot showed that: compared with the negative control group, the cell apoptosis rate was significantly lower than that in the negative control group. Overexpression of miR-497 could decrease the expression of Bcl-2 in EJ cells and increase the expression of Caspase-3. Conclusion miR-497 can inhibit the proliferation and promote the apoptosis of EJ cells, which may be related to the expression of Bcl-2 and Caspase-3.
【作者单位】：南方医科大学研究生学院;南方医科大学附属广州军区广州总医院泌尿外科;南方医科大学附属广州军区广州总医院妇产科;
【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)
【分类号】：R737.14

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