miR-1231通过靶向EGFR/PI3K/AKT通路抑制脑膜瘤细胞增殖研究

Research on MiR-1231 Inhibiting 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(P<0.05),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显著抑制脑膜瘤细胞增殖和抑制EGFR/PI3K/AKT通路(P<0.05);与anti-miR-NC组相比,anti-miR-1231显著促进脑膜瘤细胞增殖和激活EGFR/PI3K/AKT通路(P<0.05);与miR-NC+EGFR-NC组相比,miR-1231+EGFR-NC显著抑制脑膜瘤细胞EGFR/PI3K/AKT通路(P<0.05),显著下调细胞增殖能力(P<0.05);与miR-NC+EGFR-NC组相比,miR-NC+EGFR显著激活脑膜瘤细胞PI3K/AKT通路(P<0.05),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 blot was used to detect EGFR protein expression,PI3K and AKT phosphorylation levels;CCK-8 was used to detect meningioma cell viability;colony formation assay and EdU assay was used to detect meningioma cell proliferation.Results:Compared with normal meningeal cells,the expression of miR-1231 in meningiomas cells was significantly down-regulated(P<0.05),and the expression of EGFR was significantly up-regulated(P<0.05).EGFR was confirmed as a target gene of miR-1231.Compared with miR-NC group,miR-1231 significantly inhibited meningiomas cell proliferation and inhibited EGFR/PI3K/AKT pathway(P<0.05).Compared with anti-miR-NC group,anti-miR-1231 significantly promotes meningioma cell proliferation and activates the EGFR/PI3K/AKT pathway(P<0.05).Compared with miR-NC+EGFR-NC group,miR-1231+EGFR-NC significantly inhibits meningiomas cells EGFR/PI3K/AKT pathway(P<0.05),significantly reduced cell proliferation(P<0.05).Compared with miR-NC+EGFR-NC group,miR-NC+EGFR significantly activated meningiomas cells PI3K/AKT pathway(P<0.05)),significantly increased cell proliferation ability(P<0.05),and 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.