摘要
目的探究长链非编码RNA(LncRNA)LINC00667对类风湿关节炎成纤维样滑膜细胞(RA-FLS)增殖、迁移、侵袭的影响及其机制。方法复制类风湿关节炎(RA)模型大鼠,SPF级Wistar大鼠分离踝关节滑膜组织,用实时荧光定量聚合酶链反应(qRT-PCR)检测RA-FLS中LINC00667和miR-19a-3p的表达。在RA-FLS中分别转染si-NC(si-NC组)、si-LINC00667(si-LINC00667组)、miR-NC(miR-NC组)、miR-19a-3p模拟物(miR-19a-3p组)、si-LINC00667与anti-miR-NC(si-LINC00667+anti-miR-NC组)、si-LINC00667与anti-miR-19a-3p(si-LINC00667+anti-miR-19a-3p组)。CCK-8法检测各组转染后RA-FLS的增殖抑制率。Transwell法检测RA-FLS迁移、侵袭。Westernblotting检测E钙黏蛋白(Ecadherin)、N钙黏蛋白(N-cadherin)表达。双萤光素酶报告验证LINC00667和miR-19a-3p的靶向关系。结果RA模型组滑膜组织中LINC00667表达量约为正常组的270%,miR-19a-3p表达量约为正常组的36%(P<0.05)。转染成功后,si-LINC00667组RA-FLS的增殖抑制率、E-cadherin蛋白相对表达量高于si-NC组(P<0.05),si-LINC00667组迁移、侵袭细胞数、N-cadherin蛋白相对表达量低于si-NC组(P<0.05)。LINC00667靶向miR-19a-3p,miR-19a-3p组RA-FLS的增殖抑制率、E-cadherin蛋白相对表达量高于miR-NC组(P<0.05),miR-19a-3p组迁移、侵袭细胞数、N-cadherin蛋白相对表达量低于miR-NC组(P<0.05)。si-LINC00667+anti-miR-19a-3p组RA-FLS的增殖抑制率、E-cadherin蛋白相对表达量低于si-LINC00667+anti-miR-NC组(P<0.05),si-LINC00667+anti-miR-19a-3p组迁移、侵袭细胞数、Ncadherin蛋白相对表达量高于si-LINC00667+anti-miR-NC组(P<0.05)。结论抑制miR-19a-3p通过靶向LINC00667,抑制RA-FLS的增殖、迁移、侵袭,LINC00667或可用作诊断和治疗RA的靶点。
Objective To explore the effects and mechanisms of long chain non-coding RNA(lncRNA)LINC00667 on the proliferation,migration and invasion of rheumatoid arthritis synovial fibroblasts(RA-FLS).Methods The model was established in SPF Wistar rats,the synovial tissue of ankle joint was isolated,and the expression of LINC00667 and miR-19a-3p in RA-FLS was detected by quantitative real-time polymerase chain reaction(qRT-PCR).Transfect si-NC,si-LINC00667,miR-NC,miR-19a-3p simulant,si-LINC00667 and anti-miR NC,si-LINC00667 and anti-miR-19a-3p in RA-FLS,and divide them into si-NC group,si-LINC00667 group,miR NC group,miR-19a-3p group,si-LINC00667+anti-miR-NC group and si-LINC00667+anti-miR-19a-3p group.CCK-8 method was used to detect the proliferation inhibition rate of RA-FLS after different transfections,Transwell method was used to detect cell migration and invasion,and Western blotting was used to detect the expression of E cadherin and N-cadherin.Double luciferase report verified the targeting relationship between LINC00667 and miR 19a-3p.Results The expression of LINC00667 in synovial tissue of RA model group was 270%of that of normal group,and the expression of miR-19a-3p was 36%that of normal group(P<0.05).After successful transfection,the inhibition rate of RA-FLS and the expression of E-cadherin protein in the si LINC00667 group were higher than those in the si-NC group,and the number of migration and invasion cells and the expression of N-cadherin protein were lower than those in the si-NC group(P<0.05).LINC00667 targeted miR-19a-3p.The inhibition rate of RA FLS and the expression of E-cadherin protein in miR-19a-3p group were higher than those in miR-NC group,while the number of migration and invasion cells and the expression of N-cadherin protein in miR-19a-3p group were lower than those in miR-NC group(P<0.05).The inhibition rate of RA-FLS and the expression of E-cadherin protein in si-LINC00667+anti-miR-19a-3p group were lower than those in si-LINC00667+anti-miR-NC group,while the number of migration and invasion cells and the expression of N-cadherin protein in si-LINC00667+anti miR-NC group were higher than those in si-LINC00667+anti-miR-NC group(P<0.05).Conclusion LINC00667 can promote the proliferation,migration and invasion of RA-FLS by targeting miR-19a-3p.
作者
孟庆良
马俊福
高青杰
崔家康
Meng Qing-liang;Ma Jun-fu;Gao Qing-jie;Cui Jia-kang(Henan Hospital of Traditional Chinese Medicine,Zhengzhou,Henan 450002,China;Henan University of Traditional Chinese Medicine,Zhengzhou,Henan 450046,China)
出处
《中国现代医学杂志》
CAS
北大核心
2023年第15期38-45,共8页
China Journal of Modern Medicine
基金
国家自然科学基金面上项目(No:81874456)
河南省中医药科学研究重大专项课题(No:20-21ZYZD05)
河南省自然科学基金青年项目(No:232300420270)
河南省中医管理局国家中医临床研究基地科研课题(No:2021JDZX2079)
河南省中医院博士基金项目(No:2021BSJJ02)。
