LncRNA LUCAT1靶向调控miR-937-5p/MMP13轴对非小细胞肺癌细胞增殖、迁移能力的影响

LncRNA LUCAT1 promotes the proliferation and migration of non-small cell lung cancer cells by targeting the miR-937-5p/MMP13 axis

目的 探究长链非编码核糖核酸(LncRNA)肺癌相关转录物1(LUCAT1)靶向调控微小RNA-937-5p(miR-937-5p)/基质金属蛋白酶13(MMP-13)轴对非小细胞肺癌(NSCLC)细胞增殖、迁移的影响。方法 采用定量实时聚合酶链反应(qRT-PCR)检测NSCLC组织和细胞中LUCAT1、miR-937-5p和MMP-13 mRNA相对表达水平。将A549细胞分为Control组、sh-NC组、sh-LUCAT1组、sh-LUCAT1+anti-miR-937-5P组、sh-LUCAT1+OE-MMP-13组。并进行相应转染处理后,qRT-PCR和蛋白印迹检测转染效率,CCK-8和BrdU检测细胞活力和增殖;Transwell检测细胞迁移。双荧光素酶用于验证LUCAT1、miR-937-5p和MMP-13之间关系。异种移植肿瘤实验用于证实LUCAT1对肿瘤生长的影响,分为sh-NC组和sh-LUCAT1组。结果 LUCAT1和MMP-13 mRNA在NSCLC组织和细胞系中高表达,而miR-937-5p表达下调(P<0.05)。敲低LUCAT1在体外抑制A549细胞增殖和迁移,并在体内抑制肿瘤生长(P<0.05)。miR-937-5p被预测并被鉴定为LUCAT1的直接靶标,MMP-13被预测并被鉴定为miR-937-5p的靶基因,LUCAT1可通过miR-937-5p调控MMP-13表达(P<0.05)。抑制miR-937-5p或过表达MMP-13可部分逆转敲低LUCAT1在体外对细胞增殖和迁移的抑制作用(P<0.05)。结论 敲低LUCAT1通过靶向miR-937-5p间接下调MMP-13表达,抑制NSCLC细胞增殖和迁移。

Objective This study aims to explore the regulatory effect of long non-coding ribonucleic acid(lncRNA)lung cancer-associated transcript 1(LUCAT1)on the proliferation and migration of non-small cell lung cancer(NSCLC)cells by targeting the microRNA-937-5p(miR-937-5p)/matrix metalloproteinase 13(MMP-13)axis.Methods The mRNA levels of LUCAT1,miR-937-5p and MMP-13 in NSCLC tissues and cells were measured by quantitative real-time polymerase chain reaction(qRT-PCR).A549 cells were treated with blanck control,or transfected with sh-NC,sh-LUCAT1,sh-LUCAT1+anti-miR-937-5p or sh-LUCAT1+OE-MMP-13.Transfection efficacy was tested by qRT-PCR and Western blot.Cell viability and proliferation were detected by CCK-8 and BrdU assay.Cell migration was detected by Transwell assay.Dual luciferase reporter assay was performed to verify the relationship between LUCAT1,miR-937-5p and MMP-13.A xenograft tumor model was created to demonstrate the role of LUCAT1 in in vivo tumor growth,sh-NC group and sh-LUCAT1 group were allocated.Results The mRNA levels of LUCAT1 and MMP-13 were significantly upregulated,while miR-937-5p was significantly downregulated in NSCLC tissues and cell lines(P<0.05).Knockdown of LUCAT1 inhibited proliferation and migration of A549 cells in vitro,and inhibited tumor growth in vivo(P<0.05).MiR-937-5p was identified as a direct target of LUCAT1,and MMP-13 was of the downstream target of miR-937-5p.LUCAT1 could regulate MMP-13 expression through miR-937-5p(P<0.05).Kncokdown of miR-937-5p or overexpression of MMP-13 partially reversed the inhibitory effects of knockdown of LUCAT1 on cell proliferation and migration in vitro(P<0.05).Conclusion Knockdown of LUCAT1 inhibits the proliferation and migration of NSCLC by targeting miR-937-5p and thus downregulated MMP-13.