南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (1): 141-145.doi: 10.12122/j.issn.1673-4254.2021.01.21

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LncRNA MEG3通过抑制HIF1α的表达降低胶质瘤U251细胞的增殖与侵袭能力

罗奇志，张 帆，李 威，王 芳，邬力祥，黄柏胜   

  • 出版日期:2021-01-26 发布日期:2021-01-25

Overexpression of lncRNA MEG3 inhibits proliferation and invasion of glioblastoma U251 cells in vitro by suppressing HIF1α expression

  • Online:2021-01-26 Published:2021-01-25

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摘要：

目的 探讨lncRNA MEG3过表达通过调控HIF1α对胶质瘤U251细胞增殖和侵袭的影响。方法 采用荧光实时定量PCR（qRT-PCR）法检测人胚脑胶质细胞（HFGC）和胶质瘤U251细胞lncRNA MEG3和HIF1α的表达，HIF1α蛋白的表达则采用Western blot法检测；将U251细胞分为3组，6孔/组。Con组：正常培养的U251细胞；NC组：采用脂质体lipofectamine2000将pcDNA3.1空载体转染至U251细胞；pcDNA3.1-MEG3组：采用脂质体lipofectamine2000将pcDNA3.1-MEG3载体转染至U251细胞。另外，对上述3组细胞进行缺氧处理12 h。分别通过qRT-PCR法或Western blot法检测各组细胞HIF1α的表达，采用MTT法检测各组U251细胞增殖情况，Transwell侵袭实验检测各组U251细胞的侵袭。结果 U251细胞中lncRNA MEG3的表达明显低于 HFGC（P<0.05），而 HIF1α mRNA 和蛋白的表达均高于 HFGC（P<0.05）；U251 细胞中，MEG3 过表达组 HIF1α mRNA和蛋白的表达均降低（P<0.05），而且在缺氧12 h后HIF1α蛋白的表达亦降低（P<0.05）。过表达MEG3导致细胞增殖及侵袭力下降（P<0.05）。结论 lncRNA MEG3通过调控HIF1α的表达抑制U251细胞的增殖和侵袭，并可能为胶质瘤的临床治疗提供潜在的分子靶点。

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Abstract:

Objective 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 1α (HIF1α). Methods The expression of lncRNA MEG3 and HIF1α mRNA were examined in human fetal glial cells (HFGCs) and U251 cells using real-time quantitative PCR (qRT-PCR), and the expression of HIF1α protein was detected with Western blotting. U251 cells in normal culture or transfected with pcDNA3.1 vector (NC group) or pcDNA3.1-MEG3 vector via lipofectamine2000 were exposed to hypoxia for 12h, and the expressions of HIF1α 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. Results The expression of MEG3 was significantly lower and HIF1α mRNA and protein expressions were significantly higher in U251 cells than in HFGCs (P<0.05). In U251 cells, overexpression of MEG3 significantly decreased the mRNA and protein expressions of HIF1α (P<0.05). Hypoxic exposure for 12h also resulted in significantly lowered expression of HIF1α protein in U251 cells (P<0.05). Overexpression of MEG3 obviously suppressed the proliferation and invasiveness of U251 cells (P<0.05). Conclusion MEG3 overexpression inhibits the proliferation and invasion of U251 cells through suppressing the expression of HIF1α mRNA and protein, suggesting that MEG3 may serve as a potential therapeutic target for glioblastomas.

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