lncRNA CASC7靶向下调miR-181a-2-3p表达影响非小细胞肺癌细胞增殖、凋亡、迁移和侵袭  

lncRNA CASC7 affects proliferation,apoptosis,migration and invasion of nonsmall cell lung cancer cells by down-regulating miR-181a-2-3p expression

作　　者：蒙冲[1] 谢甜[1] 陈永倖[1] MENG Chong;XIE Tian;CHEN Yong-Xing(Department of Respiratory and Critical Care Medicine,Hainan Provincial People's Hospital,Haikou 570100,China)

机构地区：[1]海南省人民医院呼吸与危重症医学科,海口570100

出　　处：《中国免疫学杂志》2021年第19期2327-2331,2337,共6页Chinese Journal of Immunology

基　　金：海南省自然科学基金青年基金项目(819QN354)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)CASC7对非小细胞肺癌(NSCLC)细胞增殖、凋亡、迁移和侵袭的影响及作用机制。方法:收集2017年10月至2018年12月于本院行手术切除的39例NSCLC患者的癌组织和对应的癌旁组织,RT-qPCR检测组织中CASC7和miR-181a-2-3p表达水平,Pearson相关性分析癌组织中CASC7和miR-181a-2-3p表达的相关性。体外培养NSCLC细胞A549,双荧光素酶报告基因实验验证CASC7和miR-181a-2-3p调控关系。A549细胞分为对照组、pcDNA组、pcDNA-CASC7组、pcDNA-CASC7+miR-NC组和pcDNA-CASC7+miR-181a-2-3p组,四甲基噻唑蓝染色法(MTT)检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell检测细胞迁移和侵袭,Western blot检测细胞周期蛋白D1(CyclinD1)、活化的半胱天冬酶-3(cleaved-caspase-3)、基质金属蛋白酶2(MMP-2)和MMP-9的蛋白表达。结果:与癌旁组织比较,NSCLC患者癌组织中CASC7表达水平降低(P<0.05),miR-181a-2-3p表达水平升高(P<0.05)。NSCLC患者癌组织中CASC7与miR-181a-2-3p的表达呈负相关(r=−0.694,P<0.05)。CASC7负调控miR-181a-2-3p表达。与pcDNA组比较,pcDNA-CASC7组细胞OD值、迁移和侵袭数及Cy-clinD1、MMP-2和MMP-9蛋白表达降低(P<0.05),凋亡率和cleaved-caspase-3蛋白表达升高(P<0.05)。与pcDNA-CASC7+miR-NC组比较,pcDNA-CASC7+miR-181a-2-3p组细胞OD值、迁移和侵袭数及CyclinD1、MMP-2和MMP-9蛋白表达升高(P<0.05),凋亡率和cleaved-caspase-3蛋白表达降低(P<0.05)。而对照组与pcDNA组、pcDNA-CASC7组、pcDNA-CASC7+miR-NC组各检测指标比较差异均无统计学意义(P>0.05)。结论:NSCLC患者癌组织中CASC7表达降低,过表达CASC7可能通过下调miR181a-2-3p抑制NSCLC细胞增殖、迁移和侵袭,并诱导细胞凋亡。Objective:To investigate the effect and mechanism of long-chain non-coding RNA(lncRNA)CASC7 on cell proliferation,apoptosis,migration and invasion of non-small cell lung cancer(NSCLC).Methods:The cancer tissues and corresponding ad-jacent tissues of 39 patients with NSCLC who underwent surgical resection in our hospital from October 2017 to December 2018 were collected,and RT-qPCR detected the expression levels of CASC7 and miR-181a-2-3p,Pearson correlation analysis was used to ana-lyze the correlation between the expression of CASC7 and miR-181a-2-3p in cancer tissues.NSCLC cells A549 were cultured in vitro,and the dual luciferase reporter gene experiment verified the regulatory relationship between CASC7 and miR-181a-2-3p.A549 cells were divided into control group,pcDNA group,pcDNA-CASC7 group,pcDNA-CASC7+miR-NC group and pcDNA-CASC7+miR181a-2-3p group.Then MTT method detected cell proliferation,flow cytometry detected cell apoptosis,Transwell detected cell migra-tion and invasion,and Western blot detected the expression of CyclinD1,cleaved-caspase-3,matrix metallopro teinase-2(MMP-2)and MMP-9 protein.Results:Compared with adjacent tissues,the expression level of CASC7 in cancer tissues of NSCLC patients was re-duced(P<0.05),and miR-181a-2-3p was increased(P<0.05).There was a negative correlation between the expression of CASC7 and the expression of miR-181a-2-3p in cancer tissues of NSCLC patients(r=−0.694,P<0.05).CASC7 negatively regulated miR-181a-2-3p expression.Compared with the pcDNA group,the cell OD value,the number of migrating and invasive cells and the expression of CyclinD1,MMP-2 and MMP-9 protein in the pcDNA-CASC7 group were reduced(P<0.05),while the apoptosis rate and the expres-sion of cleared-caspase-3 protein were increased(P<0.05).Compared with the pcDNA-CASC7+miR-NC group,the cell OD value,181a-2-3p group were increased(P<0.05),while the apoptosis rate and the expression of cleared-caspase-3 protein were reduced(P<0.05).There was no statistically significant difference between the control

关 键 词：NSCLC CASC7 miR-181a-2-3p 细胞增殖 凋亡 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]