MiR-27b-3p通过下调lncRNA ST7-AS1水平调控非小细胞肺癌细胞的增殖、凋亡和侵袭

MiR-27b-3p regulates the proliferation,apoptosis,and invasion of non-small cell lung cancer cells by down-regulating lncRNA ST7-AS1

目的探究微小RNA-27b-3p(miR-27b-3p)通过调控长链非编码RNA ST7-AS1(lncRNA ST7-AS1)对非小细胞肺癌(NSCLC)细胞增殖、凋亡和侵袭的影响。方法通过qRT-PCR分析miR-27b和lncRNA ST7-AS1在人正常肺上皮BEAS-2B细胞和人NSCLC细胞系(A549、H358、H1299及NCL-H2073)中的表达情况。通过生物信息学和双荧光素酶报告基因检测分析miR-27b-3p与lncRNA ST7-AS1的靶向关系。将A549细胞分为5组:Control组、NC组、miR-27b-3p组、miR-27b-3p+VC组及miR-27b-3p+lncRNA ST7-AS1组。QRT-PCR检测miR-27b-3p和lncRNA ST7-AS1表达;MTT法检测细胞增殖活力;Annexin V-FITC/PI双染法检测细胞凋亡率;Transwell检测细胞侵袭能力。结果MiR-27b-3p在人NSCLC细胞系中表达低于人正常肺上皮细胞BEAS-2B,且在A549细胞中的表达最低(P<0.05);lncRNA ST7-AS1在人NSCLC细胞系表达水平高于BEAS-2B细胞,且在A549细胞中的表达最高(P<0.05)。生物信息学分析显示,lncRNA ST7-AS1和miR-27b-3p存在靶向结合位点;双荧光素酶报告基因检测结果显示,lncRNA ST7-AS1与miR-27b-3p存在靶向调控关系。qRT-PCR结果显示,与NC组比较,miR-27b-3p组A549细胞中miR-27b-3p水平升高,lncRNA ST7-AS1水平降低(P<0.05);与miR-27b-3p+VC组比较,miR-27b-3p+lncRNA ST7-AS1组A549细胞中miR-27b-3p水平降低,lncRNA ST7-AS1水平升高(P<0.05)。MTT、Annexin V-FITC/PI双染及Transwell结果显示,与NC组比较,miR-27b-3p组A549细胞增殖活力降低,凋亡率增高,侵袭细胞数量减少(P<0.05);与miR-27b-3p+VC组比较,miR-27b-3p+lncRNA ST7-AS1组A549细胞增殖活力增高,凋亡率降低,侵袭细胞数量增多(P<0.05)。结论miR-27b-3p通过下调lncRNA ST7-AS1水平抑制NSCLC细胞增殖和侵袭,并诱导细胞凋亡,抑制NSCLC的恶性进展。

Objective To explore the effects of microRNA-27b-3p(miR-27b-3p)on the proliferation,apoptosis,and invasion of non-small cell lung cancer(NSCLC)cells by regulating long-chain non-coding RNA ST7-AS1(lncRNA ST7-AS1).Methods The expression of miR-27b-3p and lncRNA ST7-AS1 in human normal lung epithelial BEAS-2B cells and human NSCLC cell lines(A549,H358,H1299,and NCL-H2073)were analyzed by qRT-PCR.The targeting relationship between miR-27b-3p and lncRNA ST7-AS1 was analyzed by bioinformatics and dual luciferase reporter gene detection.A549 cells were divided into five groups:Control group,NC group,miR-27b-3p group,miR-27b-3p+VC group,and miR-27b-3p+lncRNA ST7-AS1 group.The expression of miR-27b-3p and lncRNA ST7-AS1 were detected by qRT-PCR;MTT assay was used to detect the proliferation activity;Annexin V-FITC/PI double staining was used to detect the apoptosis rate;Transwell was used to detect the cell invasion.Results The expression of miR-27b-3p in human NSCLC cell lines was lower than that in human normal lung epithelial cells BEAS-2B,and the expression in A549 cells was the lowest(P<0.05);the expression levels of lncRNA ST7-AS1 in human NSCLC cell lines were higher than those in BEAS-2B cells,and were highest in A549 cells(P<0.05).Bioinformatics analysis showed that there were targeted binding sites between lncRNA ST7-AS1 and miR-27b-3p,and the dual-luciferase reporter gene assay targeted confirmed the targeting relationship between lncRNA ST7-AS1 and miR-27b-3p.The results of qRT-PCR showed that compared with the NC group,the miR-27b-3p level in A549 cells in the miR-27b-3p group was increased,while the lncRNA ST7-AS1 level was decreased(P<0.05);compared with the miR-27b-3p+VC group,the miR-27b-3p level in the miR-27b-3p+lncRNA ST7-AS1 group was decreased,while the lncRNA ST7-AS1 level was increased(P<0.05).The results of MTT,Annexin V-FITC/PI double staining,and Transwell showed that compared with the NC group,the proliferation of A549 cells in the miR-27b-3p group was decreased,the apoptosis rate was increased,and the number of invaded cells was decreased(P<0.05);compared with the miR-27b-3p+VC group,the proliferation of A549 cells in miR-27b-3p+lncRNA ST7-AS1 group was increased,the apoptosis rate was decreased,and the number of invaded cells was increased(P<0.05).Conclusion miR-27b-3p inhibits the proliferation and invasion of NSCLC cells and induces their apoptosis by down-regulating the level of lncRNA ST7-AS1,thereby inhibiting the malignant progression of NSCLC.