摘要
目的探讨Linc00891在骨肉瘤(OS)中发挥的作用并初步分析其机制。方法实时荧光定量逆转录聚合酶链反应(qPCR)检测Linc00891在不同OS细胞系U-2 OS、Saos-2、MG-63、SW 1353及人成骨细胞系hFOB 1.19中的表达,选择Saos-2和MG-63 OS细胞系转染过表达Linc00891载体。CCK-8法、平板克隆法和Transwell小室检测过表达Linc00891对人OS细胞系Saos-2和MG-63增殖、克隆形成能力和迁移能力的影响。Western blot法检测上皮间质转化相关蛋白钙黏着蛋白E(E-cad)、钙黏着蛋白N(N-cad)的表达。RNA pulldown实验检测Linc00891和miR-27a-3p在OS细胞中的结合关系。TCF/LEF报告试剂盒检测过表达Linc00891和上调miR-27a-3p对不同OS细胞系中Wnt/β-catenin信号通路的影响。结果不同OS细胞系U-2 OS、Saos-2、MG-63、SW 1353中Linc00891的表达水平显著低于正常成骨细胞系hFOB 1.19(P<0.05)。过表达Linc00891可抑制人OS细胞系Saos-2和MG-63的增殖、克隆形成能力、迁移能力及上皮间质转化(P<0.05)。Linc00891和miR-27a-3p在OS细胞中存在结合关系。过表达Linc00891能够逆转miR-27a-3p对Wnt/β-catenin信号通路的激活(P<0.05)。结论Linc00891通过竞争性结合并抑制miR-27a-3p,在OS中发挥抑癌作用。
Objective To investigate the role of Linc00891 in osteosarcoma(OS)and its mechanism.Methods The expressions of Linc00891 in different OS cell lines U-2 OS,Saos-2,MG-63,SW 1353 and human osteoblast cell line hFOB 1.19 were detected by qPCR.Saos-2 and MG-63 OS cell lines were transfected with Linc00891 vector.CCK-8 method,plate cloning method and Transwell chamber were used to detect the effects of Linc00891 overexpression on the proliferation,colony forming ability and migration ability.Western blot assay was used to detect the expressions of E-cadherin(E-cad)and N-cadherin(N-cad)related to epithelial-mesenchymal transition.RNA pulldown assay was used to detect the binding relationship between Linc00891 and miR-27a-3p in OS cells.TCF/LEF Reporter Kit was used to detect the effects of overexpression of Linc00891 and up-regulation of miR-27a-3p on Wnt/β-catenin signaling pathway in different OS cell lines.Results The expression levels of Linc00891 in different OS cell lines U-2 OS,Saos-2,MG-63 and SW 1353 were significantly lower than those in normal osteoblast cell line hFOB 1.19(P<0.05).The overexpression of Linc00891 could inhibit the proliferation,clone formation,migration and epithelial mesenchymal transition of human OS cell lines Saos-2 and MG-63(P<0.05).There was a binding relationship between Linc00891 and miR-27a-3p in OS cells.The overexpression of Linc00891 could reverse the activation of miR-27a-3p on Wnt/β-catenin signaling pathway(P<0.05).Conclusion Linc00891 plays an anti-tumor role in OS by competitively binding and inhibiting miR-27a-3p.
作者
阮成群
肖永杰
李记天
RUAN Cheng-qun;XIAO Yong-jie;LI Ji-tian(Department of Hip Injury,Luoyang Orthopedic Hospital of Henan Province(Orthopedic Hospital of Henan Province),Luoyang 471002,China;Department of Arthrology,No.988 Hospital of PLA Joint Logistics Support Force)
出处
《天津医药》
CAS
北大核心
2021年第5期449-454,共6页
Tianjin Medical Journal
基金
河南省中医药科学研究专项课题(2015ZY02063,2018ZYZD01)。
