miR-138-5p靶向缺氧诱导因子对人肾癌细胞GRC-1生长、侵袭和迁移的影响  

Effects of miR-138-5p targeting HIF-1α on cell growth,invasion and migration of renal cancer GRC-1 cells

作　　者：赵军华 周志杰 崔吉冈 倪灵凡 孟晓青 ZHAO Jun-Hua;ZHOU Zhi-Jie;CUI Ji-Gang;NI Ling-Fan;MENG Xiao-Qing(Department of Nephropathy Respiratory,Luoyang Dongfang Hospital,Luoyang 471003,China)

机构地区：[1]洛阳东方医院呼吸肾病科,洛阳471003

出　　处：《中国免疫学杂志》2019年第5期564-569,共6页Chinese Journal of Immunology

基　　金：河南省自然科学基金(No.923031300)

摘　　要：目的:本研究旨在探索miR-138-5p对人肾癌细胞GRC-1增殖、凋亡、侵袭和迁移的影响。方法:GRC-1细胞机分为5组:对照(GRC-1)组、mimic-scramble组、miR-138 mimic组、HIF-1α过表达(pc-HIF-1α)组和共转染(mimic+pc-HIF-1α)组。荧光素酶实验确定miR-138-5p与HIF-1α的靶向关系。实时定量PCR检测miR-138-5p的表达及HIF-1α的mRNA水平。蛋白印迹检测HIF-1α的蛋白水平。CCK-8检测细胞增殖。流式细胞术分析细胞凋亡。Transwell检测细胞侵袭能力。划痕实验分析细胞迁移能力。结果:荧光素酶实验表明miR-138-5p可靶向HIF-1α。miR-138 mimic组miR-138表达高于对照组(P<0. 01)。同时,miR-138 mimic组HIF-1α的mRNA水平低于对照组(P<0. 01)。miR-138 mimic组HIF-1α的蛋白水平低于对照组(P<0. 01)。与对照组相比,pc-HIF-1α组HIF-1α的蛋白水平上升(P<0. 01)。mimic+pc-HIF-1α组HIF-1α的蛋白水平低于pc-HIF-1α组(P<0. 01)。miR-138 mimic组细胞增殖倍数低于对照组(P<0. 01)。与对照组相比,pc-HIF-1α组细胞增殖倍数升高(P<0. 01)。mimic+pc-HIF-1α组细胞增殖倍数低于pc-HIF-1α组(P<0. 01)。miR-138 mimic组细胞凋亡率高于对照组(P<0. 01)。与对照组相比,pc-HIF-1α组细胞凋亡率下降(P<0. 01)。mimic+pc-HIF-1α组细胞凋亡率高于pc-HIF-1α组(P<0. 01)。miR-138 mimic组细胞侵袭和迁移能力低于对照组(P<0. 01)。与对照组相比,pc-HIF-1α组细胞侵袭和迁移能力增加(P<0. 01)。mimic+pc-HIF-1α组细胞侵袭和迁移能力低于pc-HIF-1α组(P<0. 01)。结论:miR-138-5p靶向HIF-1α可减弱人肾癌GRC-1细胞增殖、侵袭和迁移,增强细胞凋亡。Objective: This study aims to explore the effects of miR-138-5p on the cell proliferation,apoptosis,invasion and migration of renal cancer GRC-1 cells. Methods: GRC-1 cells were randomly divided into five groups,including control(GRC-1)group,mimic-scramble group,miR-138 mimic group,HIF-1α overexpression(pc-HIF-1α)group,co-transfection(mimic+pc-HIF-1α)group.The targeted relationship of miR-138-5p and HIF-1α was affirmed by luciferase assay.The expression of miR-138-5p and mRNA levels of HIF-1α were measured by qRT-PCR.The protein levels of HIF-1α were detected by Western blot.Cell proliferation was tested by CCK-8.Apoptosis was measured by flow cytometry.Cell invasion was detected by Transwell.Cell migration was tested by wound healing. Results: The luciferase assay indicated that HIF-1α was a target of miR-138-5p.The expression of miR-138 in miR-138 mimic group was higher than that of control group( P <0.01).But the mRNA and protein levels of HIF-1α in miR-138 mimic group were lower than that of control group( P <0.01).Compared with control group,the protein levels of HIF-1α in pc-HIF-1α group were increased( P <0.01).The protein levels of HIF-1α in mimic+pc-HIF-1α group were lower than that of pc-HIF-1α group( P <0.01).The proliferation folds in miR-138 mimic group were lower than that of control group( P <0.01).Compared with control group,the proliferation folds in pc-HIF-1α group were enhanced( P <0.01).The proliferation folds in mimic+pc-HIF-1α group were lower than that of pc-HIF-1α group( P <0.01).The apoptosis rate in miR-138 mimic group was higher than that of control group( P <0.01).Compared with control group,the apoptosis rate in pc-HIF-1α group was reduced( P <0.01).The apoptosis rate in mimic+pc-HIF-1α group was higher than that of pc-HIF-1α group( P <0.01).The cell invasion and migration in miR-138 mimic group were lower than that of control group( P <0.01).Compared with control group,the cell invasion and migration in pc-HIF-1α group were elevated( P <0.01).The cell invasion and

关 键 词：肾癌 miR-138-5p HIF-1Α 增殖 凋亡 侵袭 迁移 

分 类 号：R737.11[医药卫生—肿瘤]