lncRNA TTN-AS1通过miR-519d-3p/MMP2调控肺腺癌A549细胞的活力和侵袭

lncRNA TTN-AS1 regulates the viability and invasion of lung adenocar⁃cinoma A549 cell via miR-519d-3p/MMP2 pathway

目的:观察肺腺癌组织中长链非编码RNA(lncRNA)TTN反义RNA 1(TTN-AS1)的表达情况,以及沉默TTN-AS1表达对肺腺癌A549细胞活力和侵袭的影响。方法:RT-qPCR法检测32例肺腺癌和癌旁正常组织中TTN-AS1、微小RNA-519d-3p(miR-519d-3p)和基质金属蛋白酶2(MMP2)的mRNA表达水平。将未转染的A549细胞设为空白组,转染si-NC的为si-NC对照组,转染沉默TTN-AS1表达的siRNA为si-lncRNA组(n=5),采用CCK8和Transwell方法检测沉默TTN-AS1表达对A549细胞活力和侵袭的影响。使用双萤光素酶报告基因实验、RNA免疫沉淀实验、RT-qPCR和Western blot测定TTN-AS1对miR-519d-3p和miR-519d-3p对MMP2的靶向调控作用。结果:32例肺腺癌组织中TTN-AS1的表达水平显著高于对应癌旁正常组织(P<0.05)。沉默A549细胞中TTN-AS1的表达可抑制细胞的活力和侵袭。TTN-AS1可通过海绵吸附的方式负调控miR-519d-3p的表达;MMP2是miR-519d-3p的靶基因,受其负调控。过表达MMP2可部分逆转沉默TTN-AS1和过表达miR-519d-3p对A549细胞侵袭的抑制作用。结论:肺腺癌组织中lncRNA TTN-AS1呈现高表达情况,它可能通过miR-519d-3p/MMP2调控肺腺癌A549细胞的活力和侵袭能力。

AIM:To investigate the expression level of long noncoding RNA(lncRNA)TTN antisense RNA 1(TTN-AS1)in lung adenocarcinoma tissues and the effects of TTN-AS1 silencing on the viability and invasion of lung ade⁃nocarcinoma A549 cells.METHODS:RT-qPCR was used to detect the expression of TTN-AS1,microRNA-519d-3p(miR-519d-3p)and matrix metalloproteinase 2(MMP2)mRNA in 32 cases of lung adenocarcinoma and adjacent normal tissues.The untransfected A549 cells were divided into blank group,si-NC group(with si-NC transfection)and silncRNA group(with silencing of lncRNA TTN-AS1 expression),with n=5 in each group.The effects of TTN-AS1 silenc⁃ing on the viability and invasion of A549 cells were detected by CCK8 and Transwell methods.The targeting regulatory ef⁃fects of TTN-AS1 on miR-519d-3p and miR-519d-3p on MMP2 were determined by dual-luciferase reporter assay,RNA immunoprecipitation test,RT-qPCR and Western blot.RESULTS:The expression level of TTN-AS1 in 32 cases of lung adenocarcinoma tissues is notably higher than that in the adjacent normal tissues(P<0.05).Silencing of TTN-AS1 in A549 cells significantly suppressed the cell viability and invasion.TTN-AS1 negatively regulated the expression of miR-519d-3p via sponging and absorbing miR-519d-3p.MMP2 is the target gene of miR-519d-3p and can be negatively regulat⁃ed by miR-519d-3p.Overexpression of MMP2 partially reversed the inhibitory effect of TTN-AS1 silencing and miR-519d-3p overexpression on the invasion of A549 cells.CONCLUSION:The lncRNA TTN-AS1 is overexpressed in lung adeno⁃carcinoma tissues,and it regulates lung adenocarcinoma A549 cell viability and invasion via miR-519d-3p/MMP2 path⁃way.