miR-200c下调KRAS基因表达抑制膀胱癌细胞体外增殖能力  

miR-200c inhibits proliferation in bladder cancer cells through down-regulation of KRAS

作　　者：李建昌[1,2] 苏泽轩[1] 石利平[1] 陈合格[2] 柳建军[2] 刘磊[2] 

机构地区：[1]暨南大学附属第一医院泌尿外科,广州510630 [2]广东医学院附属医院泌尿外科研究室,广东湛江524001

出　　处：《第三军医大学学报》2015年第21期2142-2146,共5页Journal of Third Military Medical University

基　　金：广东省自然科学基金(2012B031800221)~~

摘　　要：目的探讨miR-200c对膀胱癌细胞体外增殖能力的作用及机制。方法慢病毒感染法建立稳定过表达和稳定低表达miR-200c膀胱癌细胞,分为对照组、稳定过表达miR-200c组(miR-200c组)、稳定低表达miR-200c组(miR-200c-inhibit组),实时定量qRT-PCR和Western blot检测不同实验组膀胱癌细胞中miR-200c、KRAS的表达量,克隆试验检测不同组膀胱癌细胞系的增殖能力。结果CCK-8试验显示对照组T24和UM-UC-3与miR-200c组分别为0.833 3±0.062 1、0.650 3±0.088 7和0.939 0±0.018 1、0.806 0±0.021 6,2组间差异有统计学意义(T24,P<0.05;UM-UC-3,P<0.01)。稳定过表达miR-200c后,膀胱癌细胞系内KRAS蛋白的表达降低(P<0.05)。结论 miR-200c在体外通过下调KRAS基因降低膀胱癌细胞的增殖能力。Objective To determine the role and underlying mechanism of miR-200c in the proliferation of bladder cancer. Methods After lentiviruses were used to stably over-express and antagonize miR-200c in bladder cancer 5637, TCC-SUP and UM-UC-3 cells, RT-qPCR and Western blotting were intended to detect the expression changes of miR-200c and KRAS. Dual luciferase reporting system was used to verify whether miR-200c regulated the 3′UTR of KRAS as target. Colony formation assay was applied to analyze the change of colony formation abilities of bladder cancer cells in different groups. Results CCK-8 assay that cell viability was 0.833 3±0.062 1 and 0.650 3±0.088 7 in the controlled and miR-200c T24 cells （P〈0.05）, and 0.939 0±0.018 1 and 0.806 0±0.021 6 in the controlled and miR-200c UM-UC-3 cells （P〈0.01）. Stably over-expression of miR-200c resulted in significant decrease in protein expression of KRAS （P〈0.05）.Conclusion miR-200c suppresses the proliferation of bladder cancer cells by down-regulating the expression of KRAS protein, which indicates that KRAS plays a vital role in the incidence and progression of bladder cancer.

关 键 词：膀胱癌 MIR-200C KRAS 增殖 

分 类 号：R394.3[医药卫生—医学遗传学] R730.23[医药卫生—基础医学]