| 褚 楚,穆 楠,顾锦涛,黄同列,舒 震,赵金康,郑朝晖,张 伟,薛晓畅.miR-10b对滑膜成纤维细胞增殖、侵袭和迁移能力的影响及分子机制[J].现代生物医学进展英文版,2018,(7):1222-1229. |
| miR-10b对滑膜成纤维细胞增殖、侵袭和迁移能力的影响及分子机制 |
| Effects and Molecular Mechanisms of miR-10b on the Proliferation, Invasion and Migration of Fibroblast-Like Synovial Cells |
| Received:December 11, 2017 Revised:December 30, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.07.005 |
| 中文关键词: 类风湿性关节炎 microRNA miR-10b 成纤维样滑膜细胞 |
| 英文关键词: Rheumatoid arthritis microRNA miR-10a Fibroblast-like synoviocytes |
| 基金项目:国家自然科学基金项目(81373201，31000406，81273279) |
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| 中文摘要: |
| 摘要 目的：探讨miR-10b对类风湿性关节滑膜成纤维细胞(RA-FLS)炎性因子分泌、增殖、侵袭和迁移的影响及其分子机制。方法：首先，分离原代培养FLS细胞并进行microarray，筛选RA和OA中差异表达的miRNA分子。然后，用real-time PCR对筛选得到的结果进行验证，进而通过生物信息学分析、细胞转染等方法明确miR-10b在FLS细胞中下调的分子机制。最后，采用MTT比色法、划痕实验和Transwell实验等检测miR-10b对其FLS细胞增殖、侵袭和迁移水平的影响。结果：与OA FLS相比，芯片筛选发现176条miRNA在RA-FLS中上调，204条下调；其中，miR-10b在RA-FLS细胞中受肿瘤坏死因子(TNF-α)等多种炎性因子以NF-κB依赖的方式进行调控；miR-10b的下游靶基因为TAK1和TLR4，通过对这两个靶基因的调控，miR-10b一方面可以促进TNF-α的分泌和NF-κB的活化入核，从而激发TNF-α→NF-κB→miR-10b→TNF-α/NF-κB环路；另一方面促进FLS表面TLR4的表达以及LPS对于FLS的刺激作用，激发LPS→NF-κB→miR-10b→TLR4环路。此外，miR-10b的下调可促进FLS细胞的增殖、侵袭和迁移。结论：miR-10b在RA-FLS细胞中显著下调，其通过参与信号环路的调节可影响FLS细胞的炎性细胞因子分泌及其增殖、侵袭和迁移。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of microRNA (miR)-10b on the inflammatory cytokine secretion, proliferation, in- vasion and migration of RA-FLS and the underlying mechanisms. Methods: Primary FLS cells were isolated for miRNA microarray to screen differentially-expressed miRNAs between RA and OA. Then, after confirming the microarray data by real-time PCR analysis, the detailed mechanisms of miR-10b down-regulation was analyzed with bioinformatics and cell transfection methods. Finally, real-time PCR, MTT assay, cell disclosure and transwell assays were used to detect the effects of miR-10b on the cytokine production, prolifera- tion, migration and invasion of FLS cells. Results: As compared with OA patients, 176 miRNAs were up-regulated, whereas 204 miRNAs were down-regulated in RA patients. Among them, miR-10b expression in RA FLS was apparently repressed by several inflammatory cy- tokines like TNF-α in a NF-κB-dependent manner. Luciferase reporter assay and real-time PCR uncovered that TAK1 and TLR4 are target genes of miR-10b. As a result, miR-10b participated in FLS function regulation via these two genes. On one hand, downregulated miR-10b greatly promoted NF-κB activation and TNF-α production through accelerating TAK1 level and then triggered TNF-α→NF-κB→miR-10b→TNF-α/NF-κB regulatory circuit. On the other, low level of miR-10b enhanced TLR4 expression on FLS and there- fore strengthened LPS stimulation. This activated another regulatory circuit LPS→NF-κB→miR-10b→TLR4. Finally, miR-10b signifi- cantly promoted FLS proliferation, invasion, migration and pro-inflammatory cytokines production through these two circuits. Conclusion: miR-10b was apparently downregulated in RA-FLS and it greatly promoted pro-inflammatory cytokine secretion, proliferation, mi- gration and invasion of FLS through two different regulatory circuits. |
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