miR-655-3p靶向KIF20A抑制人肺腺癌细胞系A549迁移及侵袭

miR-655-3p targeting KIF20A inhibits migration and invasion of lung adenocarcinoma cell line A549

目的探讨miR-655-3p靶向驱动蛋白家族成员20A(KIF20A)对肺腺癌细胞增殖、迁移及侵袭的影响。方法RT-qPCR检测人肺腺癌细胞系H460、A549、HCC-2935、H1299细胞中miR-655-3p的表达;取对数增殖期的A549细胞,分为:空白组、miR-NC组、miR-655-3p mimics组、si-NC组、si-KIF20A组、miR-655-3p mimics+OE-NC组、miR-655-3p mimics+OE-KIF20A组,RT-qPCR检测miR-655-3p表达;Western blot检测KIF20A、基质金属蛋白酶2(MMP-2)、MMP-9及RAS/丝裂原活化蛋白激酶(MAPK)通路相关蛋白表达;CCK-8法检测细胞增殖;Transwell小室法检测细胞迁移与侵袭;双荧光素酶报告基因实验验证miR-655-3p与KIF20A的关系。结果miR-655-3p在人肺腺癌细胞系中低表达,且在A549细胞中的表达量最低;过表达miR-655-3p或沉默KIF20A均能抑制A549细胞增殖、迁移、侵袭及MMP-2、MMP-9、磷酸化细胞外调节蛋白激酶1/2(p-ERK1/2)、磷酸化脂酰肌醇3激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)蛋白表达(P<0.05);miR-655-3p靶向负调控KIF20A表达,上调KIF20A可抑制过表达miR-655-3p对A549细胞增殖、迁移、侵袭及RAS/MAPK通路相关蛋白的影响。结论过表达miR-655-3p可靶向下调KIF20A,抑制肺腺癌细胞增殖、迁移与侵袭,该机制可能与抑制RAS/MAPK通路有关。

Objective To investigate the effects of miR-655-3p targeting kinesin family member 20A(KIF20A)on cell proliferation,migration and invasion of lung adenocarcinoma cells.Methods RT-qPCR was used to detect the expression of miR-655-3p in human lung adenocarcinoma cell lines H460,A549,HCC-2935,and H1299.A549 cells at logarithmic proliferation stage were divided into:blank group,miR-NC group,miR-655-3p mimics group,si-NC group,si-KIF20A group,miR-655-3p mimics+OE-NC group,miR-655-3p mimics+OE-KIF20A group.RT-qPCR was used to detect the miR-655-3p expression.Western blot was used to detect the KIF20A,matrix metalloproteinase 2(MMP-2),MMP-9,RAS/mitogen-activated protein kinas(MAPK)pathway related protein expression;CCK-8 was used to detect cell proliferation;transwell experiment was used to detect cell migration and inva-sion;dual luciferase reporter gene detection experiment was used to verify the targeting relationship between miR-655-3p and KIF20A.Results miR-655-3p was less expressed in human lung adenocarcinoma cell lines,and has the lowest expression in A549 cells.Over-expression of miR-655-3p or silencing KIF20A inhibited A549 cell proliferation,migration,invasion and MMP-2,MMP-9,phosphorylation extracellular regulatory protein kinase 1/2(p-ERK1/2),phosphoinositide 3-kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT)protein expression(P<0.05).The miR-655-3p targeted at and negatively regulated the expression of KIF20A.Up-regulation of KIF20A inhibited the effects of over-expression of miR-655-3p on the proliferation,migration,invasion and RAS/MAPK pathway related proteins of A549 cells.Conclusions Over-expression of miR-655-3p may down-regulate KIF20A and inhibit the proliferation,migration and invasion of lung adenocarcinoma cells.This mechanism may be related to the inhibition of the RAS/MAPK pathway.