抑制lncRNA LINC01503通过靶向调控miR-335-5p对肺癌细胞活力、迁移和侵袭的影响及其机制研究  

Effect of inhibiting lncRNA LINC01503 on viability,migration and invasion of lung cancer cells by targeted regulation of miR-335-5p

作　　者：杨栋勇[1] 徐源[1] 樊冀闽[1] 朱志兴[1] 张华平[1] YANG Dong-yong;XU Yuan;FAN Ji-min;ZHU Zhi-xing;ZHANG Hua-ping(Department of Respiratory and Critical Medicine,Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院呼吸与危重症医学科,福建泉州362000

出　　处：《中国病理生理杂志》2020年第4期619-627,共9页Chinese Journal of Pathophysiology

基　　金：福建省科技重大专项(No.2017YZ0001-2)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)LINC01503对肺癌细胞活力、迁移和侵袭的影响及其作用机制。方法:将人肺癌H1299细胞分为si-NC组(转染si-NC)、si-LINC01503组(转染si-LINC01503)、pcDNA组(转染pcDNA)、pcDNA-LINC01503组(转染pcDNA-LINC01503)、miR-NC组(转染miR-NC)、miR-335-5p组(转染miR-335-5p mimics)、si-LINC01503+anti-miR-NC组(共转染si-LINC01503和anti-miR-NC)、si-LINC01503+anti-miR-335-5p组(共转染si-LINC01503和anti-miR-335-5p)、miR-NC+WT-LINC01503组(共转染miR-NC和WT-LINC01503)、miR-NC+MUT-LINC01503组(共转染miR-NC和MUT-LINC01503)、miR-335-5p+WT-LINC01503组(共转染miR-335-5p和WT-LINC01503)和miR-335-5p+MUT-LINC01503组(共转染miR-335-5p和MUT-LINC01503)。采用RT-qPCR检测miR-335-5p和LINC01503的表达水平;Western blot检测蛋白表达;MTT法检测细胞活力;Transwell法检测细胞迁移和侵袭能力的变化;双萤光素酶报告基因实验验证LINC01503与miR-335-5p的靶向关系。结果:与正常肺组织相比,肺癌组织中LINC01503表达水平显著升高,miR-335-5p表达水平显著降低(P<0.05);与Ⅰ/Ⅱ期阶段相比,Ⅲ/Ⅳ期肺癌组织中LINC01503表达水平显著升高,miR-335-5p表达水平显著降低(P<0.05);LINC01503高表达的患者短期生存率显著低于LINC01503低表达的患者(P<0.05)。与正常支气管上皮细胞系BEAS-2B相比,肺癌细胞系H1299、A549和SPC-A-1中miR-335-5p的表达水平显著降低,LINC01503的表达水平显著升高(P<0.05);过表达miR-335-5p、抑制LINC01503表达均可抑制H1299细胞的活力、迁移和侵袭,抑制细胞周期蛋白D1(cyclin D1)、基质金属蛋白酶2(MMP-2)和MMP-9蛋白的表达(P<0.05)。LINC01503靶向调控miR-335-5p的表达,干扰miR-335-5p表达能逆转抑制LINC01503表达对H1299细胞活力、迁移和侵袭的抑制作用。结论:抑制lncRNA LINC01503表达可抑制肺癌细胞的活力、迁移和侵袭,其机制可能与靶向调控miR-335-5p有关。AIM:To investigate the effects of long non-coding RNA(lncRNA)LINC01503 on the viability,migration and invasion of lung cancer cells and its mechanism.METHODS:Human lung carcinoma H1299 cells were divided into si-NC group(transfected with si-NC),si-LINC01503 group(transfected with si-LINC01503),pcDNA group(transfected with pcDNA),pcDNA-LINC01503 group(transfected with pcDNA-LINC01503),miR-NC group(transfected with miR-NC),miR-335-5p group(transfected with miR-335-5p mimics),si-LINC01503+anti-miR-NC group(cotransfected with si-LINC01503 and anti-miR-NC),si-LINC01503+anti-miR-335-5p group(co-transfected with siLINC01503 and anti-miR-335-5p),miR-NC+WT-LINC01503 group(co-transfected with miR-NC and WT-LINC01503),miR-NC+MUT-LINC01503 group(co-transfected with miR-NC and MUT-LINC01503),miR-335-5p+WT-LINC01503 group(co-transfected with miR-335-5p and WT-LINC01503)and miR-335-5p+MUT-LINC01503 group(co-transfected with miR-335-5p and MUT-LINC01503).The expression of miR-335-5p and LINC01503 was detected by RT-qPCR.Western blot was used to detect protein expression.MTT assay was used to detect cell viability.Transwell assay was used to detect the migration and invasion abilities.Dual-luciferase reporter assay was used to confirm the targeted relationship between LINC01503 and miR-335-5p.RESULTS:Compared with normal tissues,the expression of LINC01503 was significantly increased in the lung cancer tissues,and the expression of miR-335-5p was significantly decreased(P<0.05).Compared with stage I/II,the expression level of LINC01503 in the lung cancer tissues of stage III/IV was significantly increased,and the expression of miR-335-5p was significantly decreased(P<0.05).The patients with high expression of LINC01503 had lower short-term survival rates than those with low expression of LINC01503(P<0.05).Compared with normal human bronchial epithelial cell line BEAS-2 B,the expression of miR-335-5p in lung cancer cell lines H1299,A549 and SPC-A-1 were significantly decreased,and the expression of LINC01503 was significantly incr

关 键 词：LINC01503 微小RNA-335-5p 肺癌 细胞活力 细胞迁移 细胞侵袭 

分 类 号：R730.23[医药卫生—肿瘤] R734.2[医药卫生—临床医学]