miR-144-3p对卵巢癌细胞增殖、迁移和侵袭的影响及机制

Effects of miR-144-3p on proliferation,migration,and invasion of ovarian cancer cells

目的探讨微小RNA-144-3p(miR-144-3p)对卵巢癌ES-2细胞增殖、迁移和侵袭的影响及作用机制。方法采用qRT-PCR法检测35例卵巢癌组织和对应癌旁组织中miR-144-3p和骨形态形成拮抗蛋白1(GREM1)mRNA表达。构建miR-144-3p高表达或GREM1敲低的ES-2细胞,采用四甲基噻唑蓝染色法(MTT)检测细胞增殖情况,Transwell实验检测细胞迁移和侵袭能力,Western blotting法检测细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶2(MMP2)和MMP9及PI3K/Akt/mTOR信号通路相关蛋白磷酸化磷脂酰肌醇3激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)和磷酸化雷帕霉素靶蛋白(p-mTOR)表达。TargetScan在线软件预测GREM1的3′UTR含有miR-144-3p的结合位点,双荧光素酶报告基因验证miR-144-3p与GREM1靶向关系。结果与癌旁组织比较,卵巢癌癌组织中miR-144-3p表达降低(P<0.05),GREM1 mRNA表达升高(P<0.05)。miR-144-3p高表达或敲低GREM1,ES-2细胞存活率、迁移和侵袭细胞数降低,CyclinD1、MMP-2、MMP-9、p-PI3K、p-Akt和p-mTOR蛋白表达降低(P均<0.05)。miR-144-3p在ES-2细胞中负调控GREM1表达,高表达GREM1能部分逆转miR-144-3p高表达对ES-2细胞增殖、迁移和侵袭及PI3K/Akt/mTOR信号通路的抑制作用。结论miR-144-3p高表达可抑制卵巢癌细胞的增殖、迁移和侵袭,其作用机制可能与下调GREM1表达及抑制PI3K/Akt/mTOR信号通路有关。

Objective To investigate the effects of miR-144-3p on the proliferation,migration,and invasion of ovarian cancer ES-2 cells and its mechanism.Methods The qRT-PCR(quantitative real-time polymerase chain reaction)was used to detect the expression levels of miR-144-3p and GREM1 mRNA in 35 cases of ovarian cancer tissues and their corresponding adjacent tissues.The ESR cells with high expression of miR-144-3p or knockdown of GREM1 were constructed.MTT was used to detect cell proliferation.Transwell was used to detect cell migration and invasion.Western blotting was used to detect the expression levels of CyclinD1,matrix metalloproteinase 2(MMP-2)and MMP-9 proteins and PI3K/AKT/mTOR signaling pathway-related protein p-PI3K,p-Akt and p-mTOR.TargetScan online software predicted that the 3'UTR of GREM1 contained a binding site for miR-144-3p.The dual luciferase reporter gene verified the targeting relationship between miR-144-3p and GREM1.Results Compared with the adjacent tissues,the expression level of miR-144-3p in the ovarian cancer tissues decreased(P<0.05),and the expression level of GREM1 mRNA increased(P<0.05).High expression of miR-144-3p or knockdown of GREM1 reduced ES-2 cell survival,migration and invasion,and decreased CyclinD1,MMP-2,MMP-9,p-PI3K,p-Akt and p-mTOR protein expression levels(all P<0.05).In the ES-2 cells,miR-144-3p negatively regulated GREM1 expression.High expression of GREM1 partially reversed the inhibitory effect of high expression of miR-144-3p on the proliferation,migration,invasion and PI3K/Akt/mTOR signaling pathway of ES-2 cells.Conclusion The high expression of miR-144-3p can inhibit the proliferation,migration and invasion of ovarian cancer cells,and its mechanism is related to down-regulation of GREM1 expression and inhibition of PI3K/Akt/mTOR signaling pathway.