Down-regulation of microRNA-142-3p inhibits the aggressive phenotypes of rheumatoid arthritis fibroblast-like synoviocytes through inhibiting nuclear factor-κB signaling (Publication with Expression of Concern. See vol. 40, 2020) (Retracted Article)
Down-regulation of microRNA-142-3p inhibits the aggressive phenotypes of rheumatoid arthritis fibroblast-like synoviocytes through inhibiting nuclear factor-κB signaling (Publication with Expression of Concern. See vol. 40, 2020) (Retracted Article)
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DOI：
10.1042/bsr20190700
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发表时间：
2019-07-08
影响因子：
4
通讯作者：
Yang, Xichao
中科院分区：
文献类型：
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作者：
Qiang, Jianhong;Lv, Tingting;Yang, Xichao
本研究旨在探讨miR-142- 3 p对类风湿关节炎（RA）人成纤维样滑膜细胞（RA HFLS）侵袭性表型的调控作用，并揭示其与核因子-κ B（NF-κ B）信号通路相关的可能机制。采用实时荧光定量PCR（qRT-PCR）和北方印迹法检测RA滑膜组织和RA-HFLS中miR-142- 3 p的表达。用miR-142- 3 p抑制剂转染RA-HFLS和/或用10 μ g/l肿瘤坏死因子α（TNF-α）处理RA-HFLS。检测细胞存活率、集落形成率、凋亡率、迁移率、侵袭力、白细胞介素6（IL-6）和基质金属蛋白酶3（MMP-3）水平。qRT-PCR检测B细胞淋巴瘤2（Bcl-2）、Bax、Bad、IL-6和MMP-3的mRNA表达。Western blot检测Bcl-2、IL-1受体相关激酶1（IRAK 1）、Toll样受体4（TLR 4）、NF-κ B p65和磷酸化NF-κ B p65（p-NF-κ B p65）的表达。通过双荧光素酶报告基因分析鉴定IRAK 1与miR-142- 3 p之间的相互作用。miR-142- 3 p在RA滑膜组织和RA-HFLS中表达上调。TNF-alpha激活RA-HFLS的侵袭性表型，包括增强增殖、迁移、侵袭和炎症，并抑制细胞凋亡。miR-142- 3 p抑制剂可显著降低TNF-α处理后RA-HFLS的细胞活力、细胞克隆数、迁移率、侵袭细胞数、IL-6和MMP-3的含量和表达，增加凋亡率、Bax和Bad的表达，降低Bcl-2的表达。miR-142- 3 p抑制剂显著逆转TNF-α处理的RA-HFLS中TNF-α诱导的IRAK 1、TLR 4和p-NF-κ B p65上调。此外，IRAK 1是miR-142- 3 p的靶点。下调miR-142- 3 p可通过抑制NF-κ B B信号通路抑制RA-HFLS的侵袭性表型。
The present study aimed to investigate the regulatory roles of miR-142-3p on the aggressive phenotypes of rheumatoid arthritis (RA) human fibroblast-like synoviocytes (RA-HFLSs), and reveal the potential mechanisms relating with nuclear factor-kappa B (NF-kappa B) signaling. miR-142-3p expression was detected in RA synovial tissues and RA-HFLSs by quantitative real-time PCR (qRT-PCR) and Northern blot analysis. RA-HFLSs were transfected with miR-142-3p inhibitor and/or treated with 10 mu g/l tumor necrosis factor alpha (TNF-alpha). The viability, colony formation, apoptosis, migration, invasion, and the levels of interleukin (IL)-6, and matrix metalloproteinase 3 (MMP-3) were detected. The mRNA expressions of B-cell lymphoma-2 (Bcl-2), Bax, Bad, IL-6, and MMP-3 were detected by qRT-PCR. Moreover, the expression of Bcl-2, IL-1 receptor-associated kinase 1 (IRAK1), Toll-like receptor 4 (TLR4), NF-kappa B p65, and phosphorylated NF-kappa B p65 (p-NF-kappa B p65) were detected by Western blot. The interaction between IRAK1 and miR-142-3p was identified by dual luciferase reporter gene assay. MiR-142-3p was up-regulated in RA synovial tissues and RA-HFLSs. TNF-alpha activated the aggressive phenotypes of RA-HFLSs, including enhanced proliferation, migration, invasion, and inflammation, and inhibited apoptosis. miR-142-3p inhibitor significantly decreased the cell viability, the number of cell clones, the migration rate, the number of invasive cells, the contents and expression of IL-6 and MMP-3, and increased the apoptosis rate and the expressions of Bax and Bad, and decreased Bcl-2 expression of TNF-alpha-treated RA-HFLSs. MiR-142-3p inhibitor significantly reversed TNF-alpha-induced up-regulation of IRAK1, TLR4, and p-NF-kappa B p65 in TNF-alpha-treated RA-HFLSs. Besides, IRAK1 was a target of miR-142-3p. The down-regulation of miR-142-3p inhibited the aggressive phenotypes of RA-HFLSs through inhibiting NF-kappa B signaling.