MicroRNA-16对类风湿关节炎患者滑膜成纤维细胞增殖、侵袭及细胞因子分泌的影响

摘要

目的：研究microRNA-16（miR-16）对类风湿关节炎（rheumatoid arthritis，RA）患者滑膜成纤维细胞（ rheumatoid arthritis synovial fibroblasts，RASFs）增殖、侵袭及细胞因子分泌的影响。方法：体外分离培养RASFs，脂质体转染化学合成的miR-16 mimic或miR-16抑制剂，分别采用MTT法、Transwell小室法和流式细胞术检测其对RASFs增殖、侵袭及凋亡的影响；RT-PCR和Western blotting检测miR-16对RASFs 基质金属蛋白酶3/13（ matrix metalloproteinase 3/13，MMP3/13）及白细胞介素1β（ interleukin 1β，IL-1β）表达的影响。结果：增殖实验结果表明miR-16可显著抑制RASFs的增殖；细胞侵袭结果表明miR-16可显著抑制RASFs的侵袭；流式细胞术检测发现miR-16对RASFs凋亡无显著影响；miR-16可下调MMP3/13及IL-1β的表达水平。结论：miR-16在RA的发生中起着重要作用，它可能通过下调MMP3/13及IL-1β的表达抑制RASFs增殖和侵袭。这为进一步研究miR-16在RA中的作用机制奠定了基础。%AIM:To investigate the effect of microRNA-16 ( miR-16) on the proliferation, invasion and cyto-kine secretion of rheumatoid arthritis ( RA) synovial fibroblasts ( RASFs) from the RA patients.METHODS: miR-16 mimic and miR-16 inhibitor were synthesized, and then Transfected into RASFs isolated from RA patients with lipo-fectamine.MTT assay, Transwell chamber and flow cytometry were used to determine the effect of miR-16 on proliferation, invasion and apoptosis of RASFs.The expression of matrix metalloproteinase 3/13 ( MMP3/13) and interleukin 1β( IL-1β) was measured by RT-PCR and Western blotting.RESULTS: The proliferation and invasion of RASFs were signifi-cantly inhibited by miR-16 mimic.The result of flow cytometry demonstrated that miR-16 had no effect on apoptosis of RASFs.Furthermore, miR-16 down-regulated the expression of MMP3/13 and IL-1β.CONCLUSION:miR-16 plays an important role in the development of RA and may inhibit the proliferation and invasion of RASFs through down-regulating the expression of MMP3/13 and IL-1β.