LncRNA LINC01224/miR-513b-5p对结肠癌细胞SW1116增殖、迁移及侵袭的影响  

Effects of lncRNA LINC01224/miR-513b-5p on proliferation,migration,and invasion of colon cancer SW1116 cells

作　　者：张兆辉[1] 王利民[1] Zhao-Hui Zhang;Li-Min Wang(Department of Hepatopancreatobiliary Surgery,Jinhua Central Hospital,Jinhua 321000,Zhejiang Province,China)

机构地区：[1]金华市中心医院肝胆胰外科,浙江省金华市321000

出　　处：《世界华人消化杂志》2021年第1期7-14,共8页World Chinese Journal of Digestology

摘　　要：背景我国结肠癌发病率与死亡率逐年上升,目前结肠癌的发病机制尚未阐明,LncRNA在结肠癌等肿瘤发生过程中可发挥重要调控作用,其主要通过竞争性结合miRNA而发挥作用,已知LncRNA LINC01224在肿瘤中可能发挥癌基因作用,但LINC01224在结肠癌形成及发展过程中的作用机制尚未阐明.目的探讨LncRNA LINC01224/miR-513b-5p对结肠癌细胞SW1116增殖、迁移、侵袭的影响及其可能作用机制.方法采用qRT-PCR法检测癌旁组织与结肠癌组织中LINC01224、miR-513b-5p的表达量;si-NC、si-LINC01224、si-LINC01224与anti-miR-NC、si-LINC01224与anti-miR-513b-5p分别转染入人结肠癌细胞SW1116;采用qRT-PCR法检测SW1116细胞中LINC01224、miR-513b-5p的表达量;采用MTT实验检测细胞存活率;应用流式细胞仪检测细胞周期;采用Transwell小室实验检测细胞迁移及侵袭;双荧光素酶报告实验检测LINC01224与miR-513b-5p的靶向关系;Western blot法检测E-cadherin、N-cadherin蛋白表达量.结果与癌旁组织比较,结肠癌组织中LINC01224的表达水平升高(P<0.05),miR-513b-5p的表达水平降低(P<0.05);与si-NC组比较,si-LINC01224组细胞存活率降低(P<0.05),G0-G1期细胞比例升高(P<0.05),S期细胞比例降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),E-cadherin蛋白水平升高(P<0.05),N-cadherin蛋白水平降低(P<0.05);双荧光素酶报告实验证实LINC01224可靶向结合miR-513b-5p;与si-LINC01224+anti-miR-NC组比较,si-LINC01224+antimiR-513b-5p组细胞存活率升高(P<0.05),G0-G1期细胞比例降低(P<0.05),S期细胞比例升高(P<0.05),迁移及侵袭细胞数增多(P<0.05),E-cadherin蛋白水平降低(P<0.05),N-cadherin蛋白水平升高(P<0.05).结论干扰LINC01224表达可通过上调miR-513b-5p而减弱结肠癌细胞增殖、迁移及侵袭能力,并可诱导细胞周期阻滞于G0-G1期.BACKGROUND The incidence and mortality of colon cancer in China are increasing year by year.At present,the pathogenesis of colon cancer has not been elucidated.Long non-coding RNAs(lncRNAs)play an important regulatory role in the occurrence of colon cancer and other tumors mainly by competitively binding microRNAs.It is known that the lncRNA LINC01224 may play an oncogenic role in tumors,but the mechanism of LINC01224 in the development and progression of colon cancer has not been elucidated.AIM To explore the effects of lncRNA LINC01224/miR-513b-5p on the proliferation,migration,and invasion of colon cancer SW1116 cells and the possible mechanism involved.METHODS qRT-PCR was used to detect the expression of LINC01224 and miR-513b-5p in colon cancer and tumor adjacent tissues.si-NC,si-LINC01224,and si-LINC01224,as well as anti-miR-NC,si-LINC01224,and anti-miR-513b-5p were transfected into human colon cancer SW1116 cells.qRT-PCR was used to detect the expression of LINC01224 and miR-513b-5p in SW1116 cells.MTT assay was used to detect the cell survival rate.Flow cytometry was used to detect the cell cycle.Transwell assay was used to detect cell migration and invasion.The dual luciferase reporter assay was used to detect the targeting relationship between LINC01224 and miR-513b-5p.Western blot method was used to detect the expression of E-cadherin and N-cadherin proteins.RESULTS Compared with adjacent tissues,the expression level of LINC01224 in colon cancer tissues was increased(P<0.05),and the expression level of miR-513b-5p was decreased(P<0.05).Compared with the si-NC group,the survival rate of cells in the si-LINC01224 group was reduced(P<0.05),the proportion of cells in the G0-G1 phase was increased(P<0.05),the proportion of cells in the S phase was reduced(P<0.05),the numbers of migrating and invasive cells were decreased(P<0.05),the protein level of E-cadherin was increased(P<0.05),and the protein level of N-cadherin was decreased(P<0.05).The dual luciferase reporter assay confirmed that LINC01224 could tar

关 键 词：LncRNA LINC01224 miR-513b-5p 结肠癌 增殖 迁移 侵袭 

分 类 号：R735.35[医药卫生—肿瘤]