■本研究旨在探讨NONHSAT042241对类风湿性关节炎成纤维细胞样滑膜细胞(RA-FLS)功能的影响及其潜在机制。
■用NONHSAT042241过表达和NONHSAT042241敲低慢病毒处理RA-FLS。细胞计数试剂盒-8(CCK-8)测定,集落形成试验,流式细胞术,Transwell分析,western-blot,ELISA,qRT-PCR检测细胞增殖的变化,凋亡,入侵,炎性细胞因子和基质金属蛋白酶(MMPs)的分泌。荧光原位杂交(FISH)分析,RNA下拉法,用质谱(MS)和RNA免疫沉淀(RIP)法寻找与NONHSAT042241结合的靶蛋白,用western-blot法检测Wnt/β-catenin信号通路相关蛋白的表达。
■NONHSAT042241过表达抑制RA-FLS的增殖(p<0.05),入侵,分泌促炎因子(IL-1和IL-6)和MMPs(MMP-1和MMP-3)(p<0.05),并升高促凋亡因子(Bax和calvedcaspase3)的水平,而NONHSAT042241敲除具有相反的效果。NONHSAT042241可以直接结合hnRNPD,并下调β-catenin的表达(p<0.05),p-GSK-3β(p<0.05),细胞周期蛋白D1(p<0.05),PCNA(p<0.05),从而减少了细胞增殖。
■NONHSAT042241可能抑制FLS介导的类风湿滑膜增殖,炎症和侵略。潜在的机制可能是NONHSAT042241抑制Wnt/β-连环蛋白信号传导的活性。
UNASSIGNED: This study aims to explore the effect of NONHSAT042241 on the function of rheumatoid arthritis -fibroblast-like synoviocyte (RA-FLS) and the underlying mechanisms.
UNASSIGNED: RA-FLS was treated with NONHSAT042241 overexpression and NONHSAT042241 knockdown lentiviruses. Cell counting kit-8 (CCK-8) assay, colony formation assay, flow cytometry, Transwell assay, western-blot, ELISA, and qRT-PCR were used to measure the changes of cell proliferation, apoptosis, invasion, secretion of inflammatory cytokines and matrix metalloproteinases (MMPs). Fluorescent in situ hybridization (FISH) assay, RNA pull-down assay, mass spectrometry (MS) and RNA immunoprecipitation (RIP) were used to find the target proteins that bond to NONHSAT042241, and western-blot was used to detect the expression of related proteins of Wnt/β-catenin signaling pathway.
UNASSIGNED: Overexpression of NONHSAT042241 inhibited the proliferation of RA-FLS (p < 0.05), invasion, secretion of pro-inflammatory factors (IL-1and IL-6) and MMPs (MMP-1 and MMP-3) (p < 0.05), and elevated the level of pro-apoptotic factors (Bax and cleaved caspase3), while NONHSAT042241 knockdown had the opposite effect. NONHSAT042241 can directly bind to hnRNP D, and down-regulated the expression of β-catenin (p < 0.05), p-GSK-3β (p < 0.05), Cyclin D1 (p < 0.05), PCNA (p < 0.05), and thus reduced the cell proliferation.
UNASSIGNED: NONHSAT042241 may inhibit FLS-mediated rheumatoid synovial proliferation, inflammation and aggression. The underlying mechanisms may be that NONHSAT042241 inhibits the activity of Wnt/β-catenin signaling.