miR-1231通过靶向EGFR/PI3K/AKT通路抑制脑膜瘤细胞增殖

Inhibitory effect of miR-1231 on meningioma cell proliferation by targeting EGFR/PI3K/AKT pathway

目的探究miR-1231通过靶向EGFR/PI3K/AKT通路对脑膜瘤细胞增殖的影响。方法生物信息学软件分析预测并通过荧光素酶报告基因实验验证miR-1231与EGFR之间的靶向结合作用,RT-PCR检测脑膜瘤细胞miR-1231表达和EGFR mRNA表达,Western blot检测EGFR蛋白表达、PI3K和AKT磷酸化水平,CCK-8检测脑膜瘤细胞活力,集落形成实验和EdU实验检测脑膜瘤细胞增殖。结果与正常脑膜细胞相比,脑膜瘤细胞中miR-1231表达显著下调,而EGFR表达显著上调(P<0.05)。生物信息学软件分析预测并通过荧光素酶报告基因证明EGFR为miR-1231的靶基因。与miR-NC组相比,miR-1231组显著抑制脑膜瘤细胞增殖、抑制EGFR/PI3K/AKT通路(P<0.05);与anti-miR-NC组相比,anti-miR-1231组显著促进脑膜瘤细胞增殖、激活EGFR/PI3K/AKT通路(P<0.05);与miR-NC+EGFR-NC组相比,miR-1231+EGFR-NC组显著抑制脑膜瘤细胞EGFR/PI3K/AKT通路,并显著下调细胞增殖能力(P<0.05);与miR-NC+EGFR-NC组相比,miR-NC+EGFR组显著激活脑膜瘤细胞PI3K/AKT通路,并显著上调细胞增殖能力(P<0.05),且能逆转miR-1231+EGFR-NC对脑膜瘤细胞PI3K/AKT通路和细胞增殖能力的抑制作用。结论miR-1231通过靶向EGFR调节PI3K/AKT通路抑制脑膜瘤细胞增殖。

Objective To investigate the effect of miR-1231 on meningioma cell proliferation by targeting the EGFR/PI3K/AKT pathway.Methods Bioinformatics software and luciferase reporter gene experiments were used to analyze and verify the targeted binding between miR-1231 and EGFR.RT-PCR was used to detect miR-1231 expression and EGFR mRNA expression in meningiomas cells.Western blotting was used to detect EGFR protein expression and phosphorylation levels of PI3K and AKT.CCK-8 was used to detect meningioma cell viability.Colony formation assay and EdU assay were used to detect meningioma cell proliferation.Results Compared with normal meningeal cells,the expression of miR-1231 in meningiomas cells was significantly down-regulated,while the expression of EGFR was significantly up-regulated(P<0.05).EGFR was confirmed as a target gene of miR-1231 forecasting and verifying by bioinformatics software and luciferase reporter gene experiments.Compared with miR-NC group,miR-1231 group significantly inhibited meningioma cell proliferation and EGFR/PI3K/AKT pathway(P<0.05).Compared with anti-miR-NC group,anti-miR-1231 group significantly promoted meningioma cell proliferation and activated the EGFR/PI3K/AKT pathway(P<0.05).Compared with miR-NC+EGFR-NC group,miR-1231+EGFR-NC group significantly inhibited EGFR/PI3K/AKT pathway of meningiomas cells,and significantly reduced cell proliferation(P<0.05).Compared with miR-NC+EGFR-NC group,miR-NC+EGFR group significantly activated PI3K/AKT pathway of meningioma cells,and significantly increased cell proliferation ability(P<0.05).Furthmore,it could significantly reverse the inhibitory effects of miR-1231+EGFR-NC on PI3K/AKT pathway and cell proliferation ability of meningiomas cells.Conclusion miR-1231 can inhibit the proliferation of meningioma cells by targeting the EGFR to regulate the PI3K/AKT pathway.