lncR FOXD2-AS1调控miR-145-5p对非小细胞肺癌细胞生物学特性的影响

Effect of lncR-FOXD2-AS1 regulating miR-145-5p on biological characteristics of non-small-cell lung cancer cells

目的 探讨长链非编码RNA(lncR)FOXD2-AS1调控微小RNA(miR)-145-5p对非小细胞肺癌(NSCLC)细胞增殖、迁移、侵袭和上皮-间质转化(EMT)等生物学特性的影响。方法 体外培养人NSCLC细胞系A549、H1299、SK-MES-1及人正常肺支气管上皮细胞系16HBE,应用RT-qPCR检测各细胞系中lncR-FOXD2-AS1和miR-145-5p表达。选取lncR-FOXD2-AS1相对高表达的SK-MES-1作为敲低实验的研究对象,将SK-MES-1细胞按照转染处理的不同分为NC组(转染si-NC)和si-FOXD2-AS1组(转染si-FOXD2-AS1),转染后,RT-qPCR检测细胞中lncR-FOXD2-AS1、miR-145-5p表达,CCK-8法检测细胞增殖活性,Transwell实验检测细胞迁移、侵袭能力,Western blotting法检测细胞中E-cadherin、N-cadherin、Fibronectin蛋白表达。双荧光素酶报告基因实验验证lncR-FOXD2-AS1和miR-145-5p的相互关系。结果 与16HBE细胞相比,lncR-FOXD2-AS1在NSCLC细胞系中高表达(P<0.05),miR-145-5p在NSCLC细胞系中低表达(P均<0.05)。与NC组比较,si-FOXD2-AS1组细胞miR-145-5p表达升高(P<0.05),0、24、48 h时OD值降低(P均<0.05),迁移及侵袭穿膜细胞数减少(P均<0.05),E-cadherin蛋白表达升高(P<0.05),N-cadherin、Fibronectin蛋白表达降低(P均<0.05)。双荧光素酶实验证实miR-145-5p是lncR-FOXD2-AS1的靶基因。结论 lncR-FOXD2-AS1在NSCLC细胞系中表达升高,敲低lncR-FOXD2-AS1通过靶向调控miR-145-5p抑制NSCLC细胞的增殖、迁移、侵袭和EMT进程。

Objective To investigate the effects of long non-coding RNA(lncR)-FOXD2-AS1 regulating microRNA(miR)-145-5p on proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT) in non-small-cell lung cancer(NSCLC) cells.Methods Human NSCLC cell lines A549,H1299,SK-MES-1,and human normal pulmonary bronchial epithelial cell line 16HBE were cultured in vitro.The expression of lncR-FOXD2-AS1 and miR-145-5p in each cell line was detected using RT-qPCR.SK-MES-1 cells with relatively high expression of lncR-FOXD2-AS1 were selected as the research object for knockdown experiments.SK-MES-1 cells were divided into the NC group(transfected with siNC) and si-FOXD2-AS1 group(transfected with si-FOXD2-AS1) according to different transfection treatments.After transfection,qRT-PCR was used to detect the expression of lncR-FOXD2-AS1 and miR-145-5p in the cells,CCK-8 was used to detect cell proliferation activity,Transwell experiment was used to detect cell migration and invasion abilities,and Western blotting was used to detect the expression of E-cadherin,N-cadherin,and Fibronectin proteins in cells.The dual luciferase experiment verified the relationship between lncR-FOXD2-AS1 and miR-145-5p.Results Compared with 16HBE cells,lncR-FOXD2-AS1 was highly expressed in NSCLC cell lines(P<0.05),and miR-145-5p was low expressed in NSCLC cell lines(P<0.05).Compared with the NC group,the expression of miR-145-5p in the si-FOXD2-AS1 group increased(P<0.05),the OD values at 0,24,and 48 h decreased(all P<0.05),the number of migrating and invading transmembrane cells became smaller(P<0.05),the expression of E-cadherin protein increased(P<0.05),and the expression levels of N-cadherin and Fibronectin proteins decreased(all P<0.05).The dual luciferase experiment confirmed that miR-145-5p was the target gene of lncR-FOXD2-AS1(P<0.05).Conclusions The expression of lncR-FOXD2-AS1 is elevated in NSCLC.Knocking down lncR-FOXD2-AS1 inhibits the proliferation,migration,invasion,and EMT processes of NSCLC cells by targeting miR-145-5p.