过表达miR-299-3p抑制人肾癌细胞系增殖和迁移

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (9): 1224-1229.

过表达miR-299-3p抑制人肾癌细胞系增殖和迁移

李玖玲^1,2, 张子归^1,2, 郭莉^1,2*

1.华中科技大学同济医学院附属武汉市中心医院肾内风湿免疫科,湖北武汉 430014;
2.华中科技大学同济医学院附属武汉市中心医院分子诊断湖北省重点实验室,湖北武汉 430014

收稿日期:2019-06-24 修回日期:2019-12-04 出版日期:2020-09-05 发布日期:2020-09-04
通讯作者: ^*glwh123456@yeah.net
基金资助:
武汉卫健委科研项目(WZ19C01)

Over-expression miR-299-3p inhibits proliferation and migration of renal carcinoma cell lines

LI Jiu-ling^1,2, ZHANG Zi-gui^1,2, GUO Li^1,2*

1. Department of Rheumatology Immunology and Nephrology;
2. Key Laboratory for Molecular Diagnosis of Hubei Province, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, China

Received:2019-06-24 Revised:2019-12-04 Online:2020-09-05 Published:2020-09-04
Contact: ^*glwh123456@yeah.net

摘要/Abstract

摘要： 目的探讨miR-299-3p在肾癌细胞系增殖、迁移和侵袭的作用及机制。方法 RT-qPCR检测肾癌细胞系(786-O、769-P、ACHN和A498)和人正常肾小管上皮细胞系(HK-2)中miR-299-3p的表达;MTT法、细胞划痕实验、Transwell小室法检测miR-299-3p mimic对肾癌786-O细胞系增殖、迁移和侵袭的影响;在线生物信息学数据库对miR-299-3p的靶基因进行了预测。Western blot检测E-cadherin,N-cadherin、vimentin和MAP3K12的蛋白表达。结果 miR-299-3p在肾癌细胞系中均低表达(P＜0.05),且在786-O细胞中表达最低。过表达miR-299-3p可显著抑制肾癌细胞系786-O增殖、迁移、侵袭、体外成瘤和上皮间质转化(EMT),综合4个数据库的结果,发现预测的miR-299-3p的共同靶基为MAP3K12,荧光素酶报告基因验证了miR-299-3p与MAP3K12的靶向关系。miR-299-3p+MAP3K12共转染786-O细胞后,能够有效降低miR-299-3p mimic诱导的细胞迁移和侵袭的抑制作用。结论 miR-299-3p在肾癌细胞中的作用机制可能是通过miR-299-3p靶向下调MAP3K12表达,调控EMT途径影响肾癌细胞增殖、迁移和侵袭。

关键词: 肾癌, 786-O细胞, miR-299-3p, MAP3K12, 上皮间质转化

Abstract: Objective To explore the effect and mechanism of miR-299-3p effects on the proliferation, migration and invasion of renal cell carcinoma cell lines. Methods RT-qPCR was used to detect the expression of miR-299-3p in renal cancer cell lines (786-O, 769-P, ACHN and A498) and human normal renal epithelial cell line(HK-2). MTT assay, wound healing assay, and Transwell assay were used to detect the effects of miR-299-3p on proliferation, migration, and invasion of 786-O cells. Online bioinformatics database was used to predict the target genes of miR-299-3p. Western blot analyzed the expression of E-cadherin,N-cadherin,vimentin and MAP3K12 in 786-O cells. Results miR-299-3p was lower expression in renal cancer cell lines and the lowest expression was found in 786-O cells. Over-expression of miR-299-3p significantly inhibited the proliferation, migration, invasion, tumor xenograft and EMT in 786-O cell and four databases predicted MAP3ZK12 as the common target of miR-299-3p. Dual luciferase reporter gene verified the targeting relationship between miR-299-3p and MAP3K12. miR-299-3p+MAP3K12 effectively reversed the inhibition of miR-299-3p mimic induced cell migration and invasion in 786-O cells. Conclusions The mechanism of miR-299-3p in renal cancer cells may be explained by down-regulated MAP3K12 to affect cell proliferation, migration, invasion and EMT pathway.

Key words: renal carcinoma, 786-O cell, miR-299-3p, MAP3K12, epithelial mesenchymal transition (EMT)

中图分类号:

R737-11

李玖玲, 张子归, 郭莉. 过表达miR-299-3p抑制人肾癌细胞系增殖和迁移[J]. 基础医学与临床, 2020, 40(9): 1224-1229.

LI Jiu-ling, ZHANG Zi-gui, GUO Li. Over-expression miR-299-3p inhibits proliferation and migration of renal carcinoma cell lines[J]. Basic & Clinical Medicine, 2020, 40(9): 1224-1229.

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