miR-212-3p靶向调控NAP1L1抑制胶质瘤细胞增殖、迁移和上皮-间充质转化

miR-212-3p Targeted Regulation of NAP1L1 Inhibits Glioma Cell Proliferation,Migration and EMT

目的探究miR-212-3p在胶质瘤细胞增殖、侵袭和上皮-间充质转化(EMT)中的分子机制。方法RT-qPCR用于衡量胶质瘤细胞中miR-212-3p和NAP1L1表达,构建NC mimic、miR-212-3p mimic、oeNC和oe-NAP1L1转染至细胞中,利用CCK-8、Transwell和伤口实验评估细胞生物学行为。通过Western blot分析EMT相关标志物的蛋白表达。miR-212-3p与NAP1L1的关系通过双荧光素酶报告基因和AgO2-RIP实验进行证实。结果miR-212-3p的表达水平在胶质瘤细胞中显著下调(P<0.0001),过表达miR-212-3p可以显著降低胶质瘤细胞增殖(P<0.0001)、侵袭(P=0.0011)和迁移能力(P<0.0001),并且抑制了EMT标志物N-钙黏蛋白(N-cadherin)(P=0.000861)和波形蛋白(Vimentin)(P=0.007430)的表达,而上调了E-钙黏蛋白(Ecadherin)(P<0.0001)的表达。miR-212-3p靶向负调控NAP1L1的表达。过表达NAP1L1逆转了miR-212-3p对胶质瘤细胞增殖(P<0.0001)、迁移(P<0.0001)和EMT(P<0.0001)的抑制作用。结论miR-212-3p通过靶向负调节NAP1L1的表达抑制胶质瘤细胞增殖、迁移和EMT。

Objective To explore the molecular mechanism of miR-212-3p in glioma cell proliferation,invasion and epithelial-mesenchymal transition(EMT).Methods The expression of miR-212-3p and NAP1L1 were detected by RT-qPCR in glioma cells.NC mimic,miR-212-3p mimic,oe-NC and oe-NAP1L1 were built and transfected in cells.CCK-8,Transwell and wound healing assay were used to evaluate the cell biological behaviour.Western blot was used to detect the expression of EMT-related biomarkers.The relationship between miR-212-3p and NAP1L1 was confirmed by the dual-luciferase reporter gene and AgO2-RIP assay.Results miR-212-3p was lowly expressed in glioma cells(P<0.0001).miR-212-3p mimic significantly inhibited the glioma cell proliferation(P<0.0001),invasion(P=0.0011)and migration(P<0.0001),and reduced the expression of EMT-related biomarkers N-cadherin(P=0.000861)and Vimentin(P=0.007430),while upregulating the expression of E-cadherin(P<0.0001).miR-212-3p targeted and negatively regulated the NAP1L1 expression.Overexpression of NAP1L1 reversed the inhibitory effects of miR-212-3p on glioma cell proliferation(P<0.0001),migration(P<0.0001),and EMT(P<0.0001).Conclusion miR-212-3p inhibits the glioma cell proliferation,migration and EMT by targeting the negative regulation of NAP1L1 expression.