| 徐大伟,马鹏举,高国军.长链非编码RNA MEG3靶向miR-21对胶质瘤U251细胞增殖、侵袭和迁移的影响[J].肿瘤学杂志,2019,25(6):524-530. |
| 长链非编码RNA MEG3靶向miR-21对胶质瘤U251细胞增殖、侵袭和迁移的影响 |
| The Effect of Long Non-coding RNA MEG3 on Cell Proliferation，Migration and Invasion of Glioma Cell Line U251 via Targeting MiR-21 |
| 投稿时间：2018-08-09 |
| DOI：10.11735/j.issn.1671-170X.2019.06.B007 |
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| 中文关键词: 胶质瘤 U251 增殖 侵袭 迁移 |
| 英文关键词:glioma U251 proliferation invasion migration |
| 基金项目:河南省科技攻关计划项目(172102310503) |
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| 中文摘要: |
| 摘 要：［目的］ 探究长链非编码RNA人母系表达基因3 (maternally expressed gene 3，MEG3)对人胶质瘤细胞U251增殖、侵袭和迁移能力的影响及机制。［方法］ RT-PCR检测MEG3和miR-21在胶质瘤组织、癌旁组织中、正常星形胶质细胞NHAs和胶质瘤细胞U251中的表达；用pcDNA-MEG3 (pc-MEG3)转染U251细胞，RT-PCR检测MEG3和miR-21的表达；生物信息及荧光素酶报告实验预测并验证MEG3和miR-21的关系；MTT检测细胞增殖能力，Transwell和划痕实验检测细胞侵袭和迁移能力；免疫印迹检测增殖细胞核抗原 (proliferating cell nuclear antigen，PCNA)、基质金属蛋白酶-2(metalloproteinase-2，MMP-2) 和MMP-9的表达。［结果］ 与癌旁组织比较，MEG3在胶质瘤组织中表达水平明显降低(t=23.169，P<0.001)，miR-21水平明显升高(t=14.965，P=0.002)；与NHAs组比较，U251组细胞MEG3表达水平明显降低(t=13.145，P<0.001)，miR-21表达水平显著升高（t=12.483，P<0.001)；pcMEG3 能显著上调MEG3的表达水平并抑制miR-21表达(t=8.129，P<0.001；t=11.705，P<0.001)；miR-21 mimic能显著促进miR-21表达并能降低MEG3 野生质粒 (MEG3 wt) 的活性(t=6.460，P<0.001；t=7.742，P=0.004)；pc-MEG3能显著降低U251细胞增殖倍数和PCNA的表达水平(F=96.45，P<0.001；t=5.337，P<0.001)，miR-21 mimic能显著减弱pc-MEG3对细胞增殖及PCNA表达的抑制作用(t=7.073，P<0.001；t=4.609，P<0.001)；同时，pc-MEG3还能显著降低U251细胞的划痕闭合率和侵袭细胞数(t=5.014，P<0.001；t=10.664，P<0.001)，并抑制MMP-2和MMP-9的表达(t=3.360，P=0.007；t=3.453，P=0.006）；miR-21 mimic能明显减弱pc-MEG3对细胞侵袭、迁移及MMP-2和MMP-9表达的抑制作用(t=2.498，P=0.032；t=4.298，P=0.002；t=4.612，P<0.001；t=5.137，P<0.001)。［结论］ MEG能通过靶向miR-21减弱胶质瘤U251细胞的增殖、侵袭和迁移能力。 |
| 英文摘要: |
| Abstract：［Objective］ To investigate the effects and mechanism of long non-coding RNA maternally expressed gene 3(MEG3) on cell proliferation，invasion and migration of glioma cell line U251. ［Methods］ The levels of MEG3 and miR-21 in normal tissue，tumor tissue，human astrocyte cell NHAs and glioma cell U251 were measured by RT-PCR. U251 cells were transferred with pcDNA-MEG3(pc-MEG3)，and the expression levels of MEG3 and miR-21 were determined by RT-PCR. Bioinformatics and luciferase reporter assay were performed for predicting and verifying the relationship of MEG3 and miR-21. Cell proliferation was determined by MTT assay，invasion and migration ability were measured by Transwell and wound healing assays. The protein levels of proliferating cell nuclear antigen (PCNA)，metalloproteinase-2(MMP-2) and MMP-9 were measured by western blot. ［Results］ Compared with normal tissue，the expression of MEG3 in glioma tumor tissues was decreased significantly(t=23.169，P<0.001)，but miR-21 was increased markedly(t=14.965，P=0.002). Compared with NHAs group，the expression level of MEG3 was down-regulated in U251 group(t=13.145，P<0.001)，while expression level of miR-21 was up-regulated notably（t=12.483，P<0.001)；pcDNA-MEG3(pc-MEG3) up-regulated the expression of MEG3，but inhibited the expression of miR-21(t=8.129，P<0.001；t=11.705，P<0.001). miR-21 mimic increased the expression level of miR-21 and decreased the activity of MEG3 wild type plasmid (MEG3 wt) (t=6.460，P<0.001；t=7.742，P=0.004). The cell growth fold and the protein level of PCNA were decreased by pc-MEG3(F=96.45，P<0.001；t=5.337，P<0.001)，miR-21 mimic attenuated the effects of pc-MEG3 on cell proliferation and PCNA(t=7.073，P<0.001；t=4.609，P<0.001). Meanwhile，pc-MEG3 decreased the wound healing closure rates and invasive cells notably(t=5.014，P<0.001；t=10.664，P<0.001)，and inhibited the expression of MMP-2 and MMP-9(t=3.360，P=0.007；t=3.453，P=0.006），while miR-21 mimic alleviated the inhibitory effects of pc-MEG3 on cell invasion，migration and the expressions of MMP-2 and MMP-9(t=2.498，P=0.032；t=4.298，P=0.002；t=4.612，P<0.001；t=5.137，P<0.001). ［Conclusion］ MEG can inhibit proliferation，invasion and migration of glioma cell line U351 by targeting miR-21. |
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