摘要
目的探讨微小RNA-182-5p(miR-182-5p)对膀胱癌细胞增殖、侵袭和迁移的影响及其分子机制。方法实时荧光定量PCR(qPCR)检测正常膀胱上皮细胞(HUC)和膀胱癌细胞(T24、5637、SW780)中miR-182-5p和同源异型盒基因B7(HOXB7)表达;生物学信息预测和双荧光素酶报告基因实验分析miR-182-5p和HOXB7的靶向关系;蛋白质印迹法(Western Blot)检测过表达miR-182-5p或抑制HOXB7表达对SW780细胞中HOXB7、P53和基质金属蛋白酶-2(MMP-2)蛋白水平的影响;噻唑蓝(MTT)比色法检测细胞增殖,Transwell小室法检测细胞侵袭和迁移。共转染pcDNA-HOXB7和miR-182-5p模拟物,观察过表达HOXB7对miR-182-5p作用于膀胱癌细胞的影响。结果HUC细胞相比,miR-182-5p在膀胱癌细胞系中表达下调(P<0.05),HOXB7 mRNA和蛋白表达上调(P<0.05)。miR-182-5p在SW780细胞中的表达量最低。HOXB7是miR-182-5p的靶基因。miR-182-5p过表达可抑制细胞活力、侵袭和迁移,降低P53和MMP-2蛋白表达量,差异有统计学意义(P<0.05),与抑制HOXB7表达结果相同。过表达HOXB7可以逆转miR-182-5p对膀胱癌细胞增殖、侵袭和迁移的抑制作用。结论miR-182-5p通过靶向调控HOXB7表达抑制膀胱癌细胞增殖、侵袭和迁移。
Objective To explore the role of microRNA-182-5 p(miR-182-5 p)in cell proliferation,invasion and migration of bladder cancer cells and the underlying mechanism.Methods The miR-182-5 p and homeobox B7 gene(HOXB7)mRNA expression levels in normal bladder epithelial cells(HUC)and bladder cancer cells(T24,5637,SW780)were detected by real-time quantitative PCR(qPCR).The biological information and the dual luciferase reporter gene assay were applied to analyze the targeting relationship between miR-182-5 p and HOXB7;Western blot was used to examine the effect of overexpressing miR-182-5 p or inhibition of HOXB7 expression on HOXB7,P53 and matrix metalloproteinases 2(MMP-2)protein levels in SW780 cells.MTT assay was employed to investigate cell proliferation,and Transwell assay was adopted to observed cell invasion and migration.The pcDNA-HOXB7 and miR-182-5 p mimics were cotransfected to investigate the impacts of HOXB7 overexpression on bladder cancer cells.Results Compared with HUC cells,miR-182-5 p expression was down-regulated in bladder cancer cells while HOXB7 mRNA and protein expressions were up-regulated(P<0.05).SW780 cells got the lowest expression level of miR-182-5 p.HOXB7 was a target gene of miR-182-5 p.Besides,overexpression of miR-182-5 p,as well as inhibition expression of HOXB7,significantly suppressed cell proliferation,invasion and migration of bladder cancer cells,reduced the levels of P53 and MMP-2 proteins,and the differences were statistically significant(P<0.05).In addition,overexpression of HOXB7 reversed the inhibitory effects of miR-182-5 p on cell proliferation,invasion and migration.Conclusion miR-182-5 p inhibits cell proliferation,invasion and migration of bladder cancer cells by targeting and regulating HOXB7.
作者
肖飞
辛士永
邵长帅
张建国
高中伟
XIAO Fei;XIN Shiyong;SHAO Changshuai;ZHANG Jianguo;GAO Zhongwei(Department of Urology,the First Affiliated Hospital of Henan University of Science and Technology,Luoyang,Henan,China,471000)
出处
《分子诊断与治疗杂志》
2020年第8期1103-1107,1123,共6页
Journal of Molecular Diagnostics and Therapy
基金
河南省科技攻关计划基金资助项目(172102310588)。
