前列腺癌中miR-140-5p靶向YES1抑制细胞增殖和迁移的作用研究  

Study on the inhibitory effects of miR-140-5p targeting YES1 proto-oncogene on cell proliferation and migration in prostate cancer

作　　者：王超[1] 梅志杰 曹振学 梁玉杰 陈梦杰 张永琪 郭园园[1] 杨小淮[1] WANG Chao;MEI Zhi-jie;CAO Zhen-xue;LIANG Yu-jie;CHEN Meng-jie;ZHANG Yong-qi;GUO Yuan-yuan;YANG Xiao-huai(Department of Urology,the First Affiliated Hospital of Bengbu Medical College,Bengbu,Anhui 233004,China)

机构地区：[1]蚌埠医学院第一附属医院泌尿外科,安徽蚌埠233004

出　　处：《中华全科医学》2021年第5期731-735,共5页Chinese Journal of General Practice

基　　金：安徽省自然科学基金(1808085QH279)。

摘　　要：目的研究miR-140-5p与YES1的靶向调控关系,阐明miR-140-5p通过YES1调控前列腺癌发生发展的作用,并研究miR-140-5p在前列腺癌细胞中的潜在作用机制。方法采用实时定量PCR检测前列腺癌组织和细胞系中miR-140-5p的表达,采用CCK-8、细胞克隆实验、迁移和划痕实验测定miR-140-5p上调对前列腺癌细胞增殖和迁移的影响,用荧光素酶报告实验和蛋白印迹实验鉴定miR-140-5p的靶基因。结果癌组织中的miR-140-5p表达量相对癌旁组织明显下调,差异有统计学意义(P<0.05)。与RWPE-1细胞相比,前列腺癌细胞系中miR-140-5p的表达水平较低,差异具有统计学意义(P<0.05)。CCK-8实验转染后miR-140-5p的细胞增殖率显著下降,细胞克隆实验实验组相比对照组细胞克隆数目偏少,差异均有统计学意义(P<0.05)。划痕实验转染miR-140-5p细胞系的愈合能力明显减弱(P<0.05);Transwell小室实验转染miR-140-5p的细胞系的迁移能力下降(P<0.05);说明过表达miR-140-5p可抑制细胞的增殖和迁移。生物信息学分析和荧光素酶报告分析确定YES1为miR-140-5p的潜在靶基因。YES1在前列腺癌细胞中过表达,与miR-140-5p表达呈负相关,差异具有统计学意义(P<0.05)。结论以上实验表明miR-140-5p通过直接靶向YES1在前列腺癌发生发展中起抑制作用,提示miR-140-5p可能是一个新的靶点,用于前列腺癌的诊断和治疗。Objective To study the targeted regulatory relationship between miR-140-5p and YES1 and elucidate the role of miR-140-5p in regulating the occurrence and development of prostate cancer through YES1.The potential mechanism of action of miR-140-5p in prostate cancer cells was also determined.Methods The expression of miR-140-5p in pros-tate cancer tissues and cell lines was detected via real-time quantitative PCR.The effects of miR-140-5p up-regulation on the proliferation and migration of prostate cancer cells were investigated via cell Counting Kit-8(CCK-8)assay,cell clo-ning experiment,and migration and scratch assay.The target gene of miR-140-5p was identified via luciferase reporter assays and Western blotting.Results The expression level of miR-140-5p in cancer tissues was significantly down-regu-lated compared with that in adjacent tissues(P<0.05).The expression level of miR-140-5p was significantly lower in prostate cancer cell lines than in RWPE-1 cells(P<0.05).After transfection via CCK-8 assay,the proliferation rate of cells transfected with miR-140-5p significantly decreased,and the number of clones in the hyroid cell clone experiment group was significantly lower than that in the control group(P<0.05).The healing ability of cell lines transfected with miR-140-5p in the scratch experiments significantly decreased(P<0.05).Transwell assay revealed that the migration a-bility of cell lines transfected with miR-140-5p(P<0.05)decreased,suggesting that overexpression of miR-140-5p can inhibit cell proliferation and migration.Bioinformatics analysis and luciferase report analysis identified YES1 as the poten-tial target gene of miR-140-5p.YES1 overexpression in prostate cancer cells was negatively correlated with miR-140-5p expression,and the difference was statistically significant(P<0.05).Conclusion These experiments demonstrated that miR-140-5p can inhibit the development of prostate cancer by directly targeting YES1,suggesting that miR-140-5p may be a new target for the diagnosis and treatment of prostate

关 键 词：miR-140-5p 前列腺癌 迁移 增殖 

分 类 号：R737.25[医药卫生—肿瘤] R730.43[医药卫生—临床医学]