摘要
目的探讨长链非编码核旁斑装配转录物1(Lnc NEAT1)靶向微小RNA(miR)-506-3p对结肠癌细胞增殖和迁移的影响及其机制。方法收集河南省人民医院手术切除的结肠癌组织标本和癌旁正常组织标本各82例,采用反转录-聚合酶链反应(RT-PCR)检测结肠癌组织和癌旁组织中Lnc NEAT1的mRNA表达水平;构建miRNA-Control、miR-506-3p和短发卡RNA(shRNA)-Control、shRNA-Lnc NEAT1慢病毒结肠癌细胞株,采用生物信息学和双荧光素酶报告基因分析Lnc NEAT1和miR-506-3p的靶向关系;采用细胞计数试剂盒(CCK-8)、Transwell分别检测shRNA-Control和shRNA-Lnc NEAT1细胞的增殖、迁移能力;蛋白质印迹法(Western blot)检测shRNA-Control和shRNA-Lnc NEAT1细胞中LAMC1的蛋白表达水平。两组均数比较采用t检验,多组间比较采用单因素方差分析。结果Lnc NEAT1在结肠癌组织(1.93±0.18)中的mRNA相对表达水平显著高于癌旁组织(0.52±0.06,t=33.337,P<0.05)。过表达Lnc NEAT1-WT后,miR-506-3p细胞的相对荧光素酶活性(0.39±0.04)较miRNA-Control组显著下降(1.50±0.12,t=26.405,P<0.05)。shRNA-Lnc NEAT1细胞48 h(0.69±0.03比1.15±0.03,t=20.129,P<0.05)和72 h(0.82±0.07比1.48±0.06,t=12.651,P<0.05)的增殖活力明显低于shRNA-Control。shRNA-Lnc NEAT1细胞的迁移细胞数明显低于shRNA-Control(47.67±5.81比155.33±12.51,t=13.649,P<0.05)。shRNA-Lnc NEAT1细胞中LAMC1的蛋白表达水平较shRNA-Control显著下降(0.48±0.04比1.39±0.11,t=30.883,P<0.05)。结论Lnc NEAT1可能通过负向抑制miR-506-3p的表达,增强LAMC1的表达,从而促进结肠癌细胞的增殖和迁移。
Objective To investigate the effect of long non-coding nuclear paraspeckle assembly transcript 1(Lnc NEAT1)targeting microRNA(miR)-506-3p on the proliferation and migration of colon carcinoma cells and its mechanism.Methods 82 cases of colon carcinoma tissue and adjacent tissue were collected in our hospital.Reverse transcriptase-polymerase chain reaction(RT-PCR)was used to detect the mRNA expression of Lnc NEAT1.The miRNA-Control,miR-506-3p and short hairpin RNA(shRNA)-Control,shRNA-Lnc NEAT1 lentivirus colon carcinoma cell lines were constructed,and the bioinformatics and dual luciferase reporter genes were used to analyze the targeting relationship between Lnc NEAT1 and miR-506-3p.Cell counting kit-8(CCK-8)and Transwell was used to detect the cell proliferation and migration.Western blotting was used to detect the expression of LAMC1.The comparison between the two groups uses t test,the multi-component comparison uses one-way analysis of variance.Results The expression of Lnc NEAT1 in colon carcinoma tissues(1.93±0.18)was significantly higher than that in adjacent tissues(0.52±0.06,t=33.337,P<0.05).Compared with miRNA-Control cells(1.50±0.12,t=26.405,P<0.05),the relative luciferase activity of miR-506-3p cells was significantly decreased(0.39±0.04)after over-expressing Lnc NEAT1-WT(P<0.05).The proliferation activity of shRNA-Lnc NEAT1 cells at 48 h(0.69±0.03 vs.1.15±0.03,t=20.129,P<0.05)and 72 h(0.82±0.07 vs.1.48±0.06,t=12.651,P<0.05)was significantly lower than that of shRNA-Control.The number of migrating cells of shRNA-Lnc NEAT1 cells(47.67±5.81 vs.155.33±12.51,t=13.649,P<0.05)was significantly reduced compared with shRNA-Control.The expression of LAMC1 in shRNA-Lnc NEAT1 cells(0.48±0.04 vs.1.39±0.11,t=30.883,P<0.05)was significantly lower than that shRNA-Control.Conclusion Lnc NEAT1 may promote the proliferation and migration of colon carcinoma cells by negatively inhibiting the expression of miR-506-3p and enhancing the expression of LAMC1.
作者
温一阳
胡晓舒
杨金花
Wen Yiyang;Hu Xiaoshu;Yang Jinhua(Department of Oncology,Henan Provincial People′s Hospital,People′s Hospital of Zhengzhou University,People′s Hospital of Henan University,Zhengzhou 450003,China;Department of Pathology,People′s Hospital of Zhengzhou,People′s Hospital of Henan University of Chinese Medicine,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第10期1915-1918,共4页
Chinese Journal of Experimental Surgery
基金
河南省医学科技攻关计划项目(201602244)。
