摘要
目的探讨负载microRNA-663a(miR-663a)的慢病毒上调miR-663a对膀胱癌EJ和5637细胞系功能学的影响。方法将负载过表达miR-663a的慢病毒转染人膀胱癌EJ和5637细胞系。EJ和5637细胞系各设干扰组、阴性对照组和空白对照组,转染72h后置于荧光显微镜下观察转染效率,采用实时定量PCR法检测miR-663a的表达量,采用细胞计数试剂盒-8(CCK-8)法检测膀胱癌细胞增殖生长情况,通过细胞划痕实验检测膀胱癌细胞迁移能力,通过Transwell小室侵袭实验检测膀胱癌细胞侵袭能力。结果 EJ和5637细胞系中,干扰组的miR-663相对表达量均显著高于空白对照组和阴性对照组(P值均<0.05),空白对照组的miR-663a相对表达量与阴性对照组间的差异均无统计学意义(P值均>0.05)。EJ细胞系转染miR-663a后第4、5天,干扰组D值显著低于阴性对照组和空白对照组同时间(P值均<0.05);5637细胞系转染miR-663a后第3、4、5天,干扰组D值显著低于阴性对照组和空白对照组同时间(P值均<0.05)。EJ和5637细胞系中,干扰组细胞培养48h时的闭合率均显著低于空白对照组和阴性对照组同时间(P值均<0.05)。EJ和5637细胞系中,干扰组Transwell小室滤膜下表面细胞数均显著低于空白对照组和阴性对照组(P值均<0.05)。结论 miR-663a体外可显著抑制膀胱癌EJ和5637细胞系的细胞增殖、迁移和侵袭能力,为miR-663a可能成为膀胱癌的治疗新靶点提供了实验依据。
Objective To investigate the effect of lentivirus-mediated microRNA-663a （miR-663a） upregulation on biological behaviors in human bladder cancer cell lines EJ and 5637 in vitro. Methods Lentivirus overloading miR-663a were transfected into EJ and 5637 cell lines. The cells were either not treated （blank group）, transfected with Lentivirus empty vector （negative control group） or transfected with lentivirus-mediated- miR-663a-（Lentivirus miR-663a group）. Transfection efficiency was determined by fluorescence microscopy 72 h after transfection.Real time fluorescene quantitation polymerase chain reaction （real-time PCR） was used to determine the expressions of miR-663a. The proliferation of bladder cancer cells was determined by cell counting kit-8 （CCK-8）. The effect of miR-663a on cancer cell migration was determined by wound healing test. The invasion was detected by Transwell test. Results The expression of miR-663a in both EJ and 5637 cells in Lentivirus miR-663a group were up-regulated significantly as compared with blank and negative control groups （all P〈0.05）. There was no significant difference in the expression of miR-663a between blank group and negative control group （both P〈0.05）. The EJ cell proliferation of Lentivirus miR-663a group were significantly suppressed in CCK-8 as compared with blank and negative control groups on day 4 and 5 after transfection （all P〈0.05）. The 5637 cell proliferation of Lentivirus miR-663a group were significantly suppressed as compared with blank and negative control groups on day 3, 4 and 5 after transfection （all P〈0.05）. In both cell lines, the rate of cell migration in Lentivirus miR-663a group were significantly lower than those in blank and negative control groups 48 h after culture （all P〈0.05）, and the invasive cells that migrated through the chamber in Lentivirus miR-663a group were significantly decreased as compared with the other two groups （all P〈0.05）. Conclusion MiR-663a can effectively suppress the proliferation, migration and invasion of human bladder cancer cell lines EJ and 5637 in vitro, miR-663a may serve as a therapeutic target for bladder cancer.
出处
《上海医学》
CAS
CSCD
北大核心
2015年第5期426-429,F0003,共5页
Shanghai Medical Journal
