miR-142-5p对人上皮性卵巢癌细胞增殖和凋亡的影响

目的:探讨miR-142-5p靶向PTEN-PI3K/Akt信号调控人上皮性卵巢癌SKOV3细胞增殖和凋亡的具体机制。方法:Real-time PCR检测miR-142-5p在人卵巢癌组织及各卵巢癌细胞系中的表达情况。利用MTT、细胞克隆形成实验观察miR-142-5p对SKOV3细胞增殖活力的影响；Caspase-3活力检测miR-142-5p对SKOV3细胞凋亡情况；信号通路抑制剂处理及Western blot检测miR-142-5p、PTEN与PI3K/Akt信号通路的关系及PI3K/Akt信号下游基因Akt、FOXO1、cyclin D1等的表达情况；Luciferase实验验证miR-142-5p和PTEN 3' UTR的结合。结果:人上皮性卵巢癌组织及其细胞系中miR-142-5p表达显著增加（P<0.05)。与对照组相比，SKOV3细胞转染miR-142-5p mimics后活细胞数量显著增加，增殖能力显著上升，细胞凋亡下降（P<0.05)；与之相反的，转染miR-142-5p inhibitor后SKOV3细胞增殖能力下调，凋亡增加。信号通路抑制剂处理显示miR-142-5p过表达激活PI3K/Akt信号通路。Luciferase实验显示miR-142-5p与PTEN 3' UTR直接结合。与对照组相比，miR-142-5p mimics组PTEN表达显著降低，p-Akt蛋白表达则显著上调（P<0.05），同时过表达PTEN后p-Akt表达则显著降低（P<0.05）。结论:miR-142-5p通过抑制PTEN基因表达活化PI3K/Akt信号通路，促进人上皮性卵巢癌组织及SKOV3细胞增殖存活，抑制细胞凋亡。

Effects of miR-142-5p on proliferation and apoptosis of human epithelial ovarian cancer cells

Objective:To investigate the mechanism of miR-142-5p regulating proliferation and apoptosis of human epithelial ovarian cancer SKOV3 cells by targeting PTEN-PI3K/Akt signaling.Methods:miR-142-5p expression in ovarian cancer tissues and cell lines were detected by Real-time PCR.The SKOV3 cell proliferation rate was measured by MTT assay and colony formation assay and the cell apoptosis was detected by Caspase-3 activity.PI3K inhibitor treatment and Western blot proved that miR-142-5p functions associated with PI3K/Akt signaling and the protein expression of PTEN，Akt，FOXO1，cyclin D1.The Luciferase assay detected the PTEN 3' UTR binding with miR-142-5p.Results:The miR-142-5p expression was up-regulated in ovarian cancer tissues and cell lines.Compared with control，the number of living cells and the proliferative ability increased significantly，and the apoptotic rate decreased significantly after SKOV3 cells transfected with miR-142-5p mimics.In contrast，SKOV3 cells transfected with miR-142-5p inhibitor showed decreased proliferation and increased apoptosis(P<0.05).PI3K inhibitor treatment showed overexpression of miR-142-5p activited PI3K-Akt signaling pathway.The Luciferase assay result showed the PTEN 3' UTR directly binding with miR-142-5p.Compared with the control，the expression of p-Akt was significantly increased in miR-142-5p mimics group and PTEN expression was decreased，meanwhile，overexpression of PTEN significantly decreased p-Akt expression.Conclusion:miR-142-5p promote the proliferation and survival of human ovarian cancer tissues and cell lines and inhibit the cell apoptosis through down-regulated PTEN expression and activated the PI3K/Akt signaling pathway.