hsa-miR-150-5p靶向HIF1α对成胶质细胞瘤U-251MG细胞恶性生物学行为的影响  

Regulatory effect of hsa-miR-150-5p on malignant behaviors of glioblastoma U-251MG cells by targeting HIF1α

作　　者：周江 夏祥国 陈礼刚 罗鑫 阿库布千 郭张超 ZHOU Jiang, XIA Xiangguo, CHEN Ligang, LUO Xin, AKU Buqian, GUO Zhangchao(Department of Neurosurgery, Affiliated Hospital of Southwestern Medical University, Luzhou 646000, Sichuan, China)

机构地区：[1]西南医科大学附属医院神经外科,四川泸州646000

出　　处：《中国肿瘤生物治疗杂志》2018年第9期878-883,共6页Chinese Journal of Cancer Biotherapy

基　　金：四川省卫生和计划生育委员会资助课题(No.1504122)~~

摘　　要：目的:探讨hsa-miR-150-5p靶向低氧诱导因子1α(hypoxia inducibility factor 1,HIF1α)对成胶质细胞瘤(glioblastoma,GBM)U-251MG细胞恶性生物学行为的影响及其作用机制。方法:q RT-PCR检测miR-150-5p及其HIF1αm RNA在U-251MG细胞中的表达水平,荧光素酶报告实验验证miR-150-5p和HIF1α之间的生物学关系及miR-150-5p和HIF1α在U-251MG细胞中的生物学功能,Western blotting检测miR-150-5p及HIF1α蛋白在U-251MG细胞中的表达,Transwell实验检测U-251MG细胞的侵袭能力,划痕实验检测U-251MG细胞的迁移能力。结果:miR-150 mimic转染U-251MG细胞后,HIF1αm RNA表达水平明显下调(P<0.01),HIF1α蛋白水平也明显下调(P<0.01)。miR-150-5p降低了wt HIF1α3’-UTR转染细胞的荧光素酶活性(P<0.05),证实miR-150-5p通过结合HIF1α的3'-UTR负调控HIF1α的表达。在U-251MG细胞中,miR-150-5p过表达可明显抑制HIF1α蛋白表达、细胞侵袭和迁移(均P<0.05)。结论:miR-150-5p通过负调控HIF1α抑制U-251MG细胞的侵袭和转移,表明miR-150-5p和HIF1α有可能是GBM潜在的治疗靶点。Objective： To explore the effect and possible mechanisms of has-miR-150-5 p targeting HIF1α to regulate malignant biological behaviors of glioblastoma（GBM） U-251 MG cells. Methods： Real-time quantitative PCR（RT-PCR） was used to detect the expression of miR-150-5 p and hypoxia inducible factor 1（HIF1α） in U-251 MG cells. Luciferase report assay was carried out to verify the biological relationship between miR-150-5 p and HIF1α and their biological functions in U-251 MG cells. The protein expressions of miR-150-5 pand HIF1α in U-251 MG cells were detected by western blotting. The ability of cell migration was detected by wound healing test and cell invasion ability was detected by transwell test. Results： After miR-150-5 p mimic transfection, the m RNA expression of HIF1α was significantly reduced in U-251 MG cells（P〈0.01）. Bioinformatics prediction and luciferase reporter assay demonstrated that miR-150-5 p down-regulated HIF1α through directly binding to HIF1α 3＇-untranslated region（3＇-UTR）（all P〈0.05）. In U-251 MG cells, miR-150-5 p over-expression significantly inhibited HIF1α expression, cell invasion and migration（all P〈0.05）. Conclusion： miR-150-5 p inhibits cell invasion and metastasis through negative regulation of HIF1α, indicating that miR-150-5 p and HIF1α were both potential therapeutic targets for glioblastoma.

关 键 词：成胶质细胞瘤 miR-150-5p 低氧诱导因子1Α 侵袭 迁移 

分 类 号：R392.11[医药卫生—免疫学] R739.41[医药卫生—基础医学]