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抑制LINC00667表达对类风湿关节炎成纤维样滑膜细胞增殖、迁移、侵袭的影响
引用本文:孟庆良,马俊福,高青杰,崔家康.抑制LINC00667表达对类风湿关节炎成纤维样滑膜细胞增殖、迁移、侵袭的影响[J].中国现代医学杂志,2023(15):38-45.
作者姓名:孟庆良  马俊福  高青杰  崔家康
作者单位:1.河南省中医院, 河南 郑州 450002;2.河南中医药大学, 河南 郑州 450046
基金项目: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)
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。Western blotting检测E钙黏蛋白(E-cadherin)、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、N-cadherin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

关 键 词:类风湿关节炎  成纤维样滑膜细胞  LINC00667  miR-19a-3p  增殖  迁移  侵袭
收稿时间:2023/1/20 0:00:00

Effects of inhibition of IC06LINC00667 expression on proliferation, migration and invasion of fibroblastic synovial cells in rheumatoid arthritis
Meng Qing-liang,Ma Jun-fu,Gao Qing-jie,Cui Jia-kang.Effects of inhibition of IC06LINC00667 expression on proliferation, migration and invasion of fibroblastic synovial cells in rheumatoid arthritis[J].China Journal of Modern Medicine,2023(15):38-45.
Authors:Meng Qing-liang  Ma Jun-fu  Gao Qing-jie  Cui Jia-kang
Institution:1.Henan Hospital of Traditional Chinese Medicine, Zhengzhou, Henan 450002, China;2.Henan University of Traditional Chinese Medicine, Zhengzhou, Henan 450046, China
Abstract: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.
Keywords:arthritis  rheumatoid  synoviocytes  fibroblast-like  LINC00667  miR-19a-3p  proliferation  migration  invasion
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