Luo Qizhi, Zhang Fan, Li Wei, Wang Fang, Wu Lixiang, Huang Baisheng
Department of Immunology, School of Basic Medical Sciences, Central South University, Changsha 410008, China.
Department of Physiology, School of Basic Medical Sciences, Central South University, Changsha 410008, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2021 Jan 30;41(1):141-145. doi: 10.12122/j.issn.1673-4254.2021.01.21.
To investigate the effects of overexpression of long noncoding RNA (lncRNA) MEG3 on the proliferation and invasion of glioblastoma U251 cells by suppressing the expression of hypoxia inducible factor 1(HIF1).
The expression of lncRNA MEG3 and HIF1 mRNA were examined in human fetal glial cells (HFGCs) and U251 cells using realtime quantitative PCR (qRT-PCR), and the expression of HIF1 protein was detected with Western blotting.U251 cells in normal culture or transfected with pcDNA3.1 vector (NC group) or pcDNA3.1-MEG3 vector lipofectamine2000 were exposed to hypoxia for 12h, and the expressions of HIF1 mRNA and protein were detected with qRT-PCR and Western blotting, respectively.MTT assay and Transwell assay were employed to examine the influence of MEG3 overexpression on the proliferation and invasion of U251 cells.
The expression of MEG3 was significantly lower and HIF1 mRNA and protein expressions were significantly higher in U251 cells than in HFGCs ( < 0.05).In U251 cells, overexpression of MEG3 significantly decreased the mRNA and protein expressions of HIF1( < 0.05).Hypoxic exposure for 12h also resulted in significantly lowered expression of HIF1 protein in U251 cells ( < 0.05).Overexpression of MEG3 obviously suppressed the proliferation and invasiveness of U251 cells ( < 0.05).
MEG3 overexpression inhibits the proliferation and invasion of U251 cells through suppressing the expression of HIF1 mRNA and protein, suggesting that MEG3 may serve as a potential therapeutic target for glioblastomas.
通过抑制缺氧诱导因子1（HIF1）的表达，研究长链非编码RNA（lncRNA）MEG3过表达对胶质母细胞瘤U251细胞增殖和侵袭的影响。
采用实时定量聚合酶链反应（qRT-PCR）检测人胎儿神经胶质细胞（HFGCs）和U251细胞中lncRNA MEG3和HIF1 mRNA的表达，并用蛋白质免疫印迹法检测HIF1蛋白的表达。将正常培养的U251细胞或用pcDNA3.1载体（NC组）或pcDNA3.1-MEG3载体经脂质体2000转染后的U251细胞置于缺氧环境中12小时，分别用qRT-PCR和蛋白质免疫印迹法检测HIF1 mRNA和蛋白的表达。采用MTT法和Transwell法检测MEG3过表达对U251细胞增殖和侵袭的影响。
U251细胞中MEG3的表达明显低于HFGCs，而HIF1 mRNA和蛋白的表达明显高于HFGCs（P<0.05）。在U251细胞中，MEG3过表达显著降低了HIF1的mRNA和蛋白表达（P<0.05）。缺氧暴露12小时也导致U251细胞中HIF1蛋白表达显著降低（P<0.05）。MEG3过表达明显抑制了U251细胞的增殖和侵袭能力（P<0.05）。
MEG3过表达通过抑制HIF1 mRNA和蛋白的表达来抑制U251细胞的增殖和侵袭，提示MEG3可能成为胶质母细胞瘤的潜在治疗靶点。
