Department of Orthopedics, Shandong Provincial Western Hospital, Jinan, China.
Int J Rheum Dis. 2020 Aug;23(8):1066-1075. doi: 10.1111/1756-185X.13901. Epub 2020 Jun 29.
To explore the roles of miR-1193/Janus kinase 3 (JAK3) axis and the potential mechanism in rheumatoid arthritis (RA).
The dysregulated genes and microRNAs (miRNAs) were screened using the datasets of GSE12021 and GSE72564, while the upstream miRNA of JAK3 was forecasted by TargetScan. Then the MH7A cells were treated with interleukin-1β (IL-1β) to induce local inflammation. Double luciferase report assay was used to estimate the association between JAK3 and miR-1193. Flow cytometry and 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assays were taken to analyze the apoptosis and proliferation of MH7A cells with IL-1β-induced inflammation, respectively. The relative expression of genes and proteins were detected by quantitative real-time polymerase chain reaction and western blot analyses. Finally, rescue experiments were employed to explore the effects of miR-1193/JAK3 axis on IL-1β-induced inflammation.
JAK3 was notably up-regulated in RA, and was targeted and negatively regulated by miR-1193 which was lowly expressed in RA tissues and IL-1β-treated cells. MiR-1193 mimic significantly inhibited while miR-1193 inhibitor promoted the proliferation of MH7A cells with IL-1β-induced inflammation. Furthermore, overexpression of JAK3 presented auxo-action while depletion of JAK3 exhibited inhibitory effect on the proliferation of MH7A cells with IL-1β-induced inflammation, which could be weakened by miR-1193 mimic and miR-1193 inhibitor, respectively. Analogously, JAK3 recovered the cell proliferation that inhibited by miR-1193 mimic and inhibited cell apoptosis enhanced by miR-1193 mimic.
Up-regulation of miR-1193 suppressed the proliferation and expedited the apoptosis of MH7A cells with IL-1β-induced inflammation through targeting JAK3, which might provide novel understanding on the mechanism underlying RA.
探讨 miR-1193/Janus 激酶 3（JAK3）轴在类风湿关节炎（RA）中的作用及潜在机制。
利用 GSE12021 和 GSE72564 数据集筛选失调基因和 microRNAs（miRNAs），利用 TargetScan 预测 JAK3 的上游 miRNA。然后用白细胞介素-1β（IL-1β）处理 MH7A 细胞诱导局部炎症。双荧光素酶报告实验评估 JAK3 与 miR-1193 之间的关联。流式细胞术和 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐（MTT）检测分别分析 IL-1β 诱导炎症对 MH7A 细胞凋亡和增殖的影响。通过定量实时聚合酶链反应和 Western blot 分析检测基因和蛋白的相对表达。最后，采用 rescue 实验探讨 miR-1193/JAK3 轴对 IL-1β 诱导炎症的影响。
JAK3 在 RA 中显著上调，在 RA 组织和 IL-1β 处理的细胞中低表达，是 miR-1193 的靶基因并受其负调控。miR-1193 模拟物显著抑制，而 miR-1193 抑制剂促进 IL-1β 诱导炎症的 MH7A 细胞增殖。此外，JAK3 过表达表现出辅助作用，而 JAK3 耗竭对 IL-1β 诱导炎症的 MH7A 细胞增殖具有抑制作用，分别用 miR-1193 模拟物和 miR-1193 抑制剂可减弱这种作用。类似地，JAK3 恢复了 miR-1193 模拟物抑制的细胞增殖，并抑制了 miR-1193 模拟物增强的细胞凋亡。
miR-1193 的上调通过靶向 JAK3 抑制 IL-1β 诱导炎症的 MH7A 细胞增殖并促进其凋亡，为 RA 发病机制提供了新的认识。
