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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[J].周江,夏祥国,陈礼刚,罗鑫,阿库布千,郭张超.中国肿瘤生物治疗杂志,2018,(9):878~883.
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    点此下载全文    点此浏览HTML全文
周江  夏祥国  陈礼刚  罗鑫  阿库布千  郭张超
西南医科大学附属医院神经外科,四川泸州646000
基金项目:四川省卫生和计划生育委员会资助课题(No.1504122)
DOI:10.3872/j.issn.1007-385X.2018.09.006
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:qRT-PCR检测miR-150-5p 及其HIF1α mRNA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,HIF1α mRNA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 HIF1α 3’-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。
关键词:成胶质细胞瘤  miR-150-5p  低氧诱导因子1α  侵袭  迁移
Regulatory effect of hsa-miR-150-5p on malignant behaviors of glioblastoma U-251MG cells by targeting HIF1α    Download Fulltext
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
Fund Project:Project supported by the Health and Family Planning Commission of Sichuan Province (No.1504122)
Abstract:
      Objective: To explore the effect and possible mechanisms of has-miR-150-5p targeting HIF1α to regulate malignant biological behaviors of glioblastoma (GBM) U-251MG cells. Methods: Real-time quantitative PCR (RT-PCR) was used to detect the expression of miR-150-5p and hypoxia inducible factor 1 (HIF1α) in U-251MG cells. Luciferase report assay was carried out to verify the biological relationship between miR-150-5p and HIF1α and their biological functions in U-251MG cells. The protein expressions of miR-150-5pand HIF1α in U-251MG 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-150-5p mimic transfection, the mRNA expression of HIF1α was significantly reduced in U-251MG cells (P<0.01). Bioinformatics prediction and luciferase reporter assay demonstrated that miR-150-5p down-regulated HIF1α through directly binding to HIF1α 3’- untranslated region (3’-UTR) (all P<0.05). In U-251MG cells, miR-150-5p over-expression significantly inhibited HIF1α expression, cell invasion and migration (all P<0.05). Conclusion: miR-150-5p inhibits cell invasion and metastasis through negative regulation of HIF1α, indicating that miR-150-5p and HIF1α were both potential therapeutic targets for glioblastoma.
Keywords:glioblastoma(GBM)  miR-150-5p  hypoxia inducible factor 1α(HIF1α)  invasion  migration
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