miR-526b-3p调控TROAP基因介导Wnt信号通路对非小细胞肺癌细胞增殖与侵袭迁移的影响  

Influence of miR-526b-3p on the proliferation,invasion,and migration of non-small cell lung cancer cells by regulating TROAP gene-mediated Wnt signaling pathway

作　　者：邓科兰[1] 刘桂霞[1] DENG Kelan;LIU Guixia(Department of Respiratory and Critical Care Medicine,Xiaogan Central Hospital,Xiaogan,Hubei 432099,China)

机构地区：[1]孝感市中心医院呼吸与危重症医学科,湖北孝感432099

出　　处：《临床肺科杂志》2023年第7期1065-1073,共9页Journal of Clinical Pulmonary Medicine

基　　金：孝感市自然科学计划项目(No.XGKJ2020010004)。

摘　　要：目的探究miR-526b-3p调控滋养素相关蛋白(TROAP)基因介导Wnt信号通路对非小细胞肺癌(NSCLC)细胞增殖与侵袭迁移的影响。方法测定人肺泡上皮细胞及NSCLC细胞系A549、HCC827、NCI-H1299中miR-526b-3p及TROAP、Wnt基因的表达水平。将体外培养的A549细胞随机分为对照组、miR-526b-3p mimic组、miR-526b-3p mimic阴性对照组、miR-526b-3p mimic+TROAP过表达组、miR-526b-3p mimic+Wnt过表达组,分组转染后,测定各组细胞增殖、凋亡、迁移、侵袭、细胞增殖相关基因(Cyclin D1)、凋亡相关基因(Bax、Caspase-3)、上皮间质转化相关基因(E-cadherin、Vimentin)、TROAP基因、Wnt基因及miR-526b-3p表达、细胞Cyclin D1、Bax、Caspase-3、E-cadherin、Vimentin、TROAP及Wnt蛋白表达、miR-526b-3p对TROAP的靶向调控作用。结果与对照组相比,miR-526b-3p mimic组细胞呈现凋亡损伤状态,细胞Edu阳性率、细胞迁移率、细胞侵袭数、细胞Cyclin D1、Vimentin、TROAP、Wnt mRNA及蛋白表达水平显著降低(P<0.05),细胞miR-526b-3p表达、Bax、Caspase-3、E-cadherin mRNA及蛋白表达水平显著升高(P<0.05);miR-526b-3p mimic阴性对照组细胞各指标无明显差异(P>0.05)。过表达TROAP或Wnt可消除miR-526b-3p mimic对细胞增殖与侵袭迁移的作用。在A549细胞中miR-526b-3p可靶向下调TROAP的表达。结论miR-526b-3p可通过靶向下调TROAP的表达而降低Wnt基因表达,进而抑制A549细胞周期相关基因表达,促进细胞凋亡,减轻上皮间质转化,最终降低NSCLC增殖与侵袭迁移活性。Objective To investigate the influence of miR-526b-3p on the proliferation,invasion and migration of non-small cell lung cancer(NSCLC)cells by regulating Trophinin-associated protein(TROAP)-mediated Wnt signaling pathway.Methods The expression of miR-526b-3p,TROAP,and Wnt genes in human alveolar epithelial cells and NSCLC cell lines A549,HCC827 and NCI-H1299 was determined respectively.The A549 cells cultured in vitro were randomly divided into the control group,the miR-526b-3p mimic group,the miR-526b-3p mimic negative control group,the miR-526b-3p mimic+TROAP overexpression group,and the miR-526b-3p mimic+Wnt overexpression group.After grouping and transfection,their cell proliferation,apoptosis,migration,invasion,mRNA expression of cell proliferation-related genes(Cyclin D1),apoptosis-related genes(Bax,Caspase-3),epithelial-mesenchymal transition-related genes(E-cadherin,Vimentin),TROAP genes,Wnt Gene and miR-526b-3p,protein expression of Cyclin D1,Bax,Caspase-3,E-cadherin,Vimentin,TROAP and Wnt in cells were measure in each group.The targeted regulation of TROAP by miR-526b-3p in A549 cells was investigated.Results Compared with the control group,the cells in the miR-526b-3p mimic group showed apoptosis damage,Edu positive rate,cell migration rate,cell invasion number,cell Cyclin D1,Vimentin,TROAP,Wnt mRNA and protein expression levels were significantly decreased(P<0.05),the miR-526b-3p expression level,Bax,Caspase-3,E-cadherin mRNA and protein expression levels in cells were significantly increased(P<0.05).There was no significant difference in each index of cells in the miR-526b-3p mimic negative control group(P>0.05).Over-expression of TROAP or Wnt could eliminate the effect of miR-526b-3p mimic on cell proliferation,invasion and migration.MiR-526b-3p was able to target down-regulation of TROAP expression in A549 cells.Conclusion MiR-526b-3p can reduce Wnt gene expression through targeted down-regulation of TROAP expression,thereby inhibiting the expression of A549 cell cycle-related genes,promoting cell

关 键 词：miR-526b-3p 滋养素相关蛋白 WNT信号 非小细胞肺癌 增殖 侵袭迁移 

分 类 号：R734.2[医药卫生—肿瘤]