MicroRNA-126 affects rheumatoid arthritis synovial fibroblast proliferation and apoptosis by targeting PIK3R2 and regulating PI3K-AKT signal pathway.
MicroRNA-126 affects rheumatoid arthritis synovial fibroblast proliferation and apoptosis by targeting PIK3R2 and regulating PI3K-AKT signal pathway.
复制标题
MicroRNA-126通过靶向PIK3R2并调节PI3K-AKT信号通路影响类风湿性关节炎滑膜成纤维细胞增殖和凋亡
DOI：
10.18632/oncotarget.12487
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发表时间：
2016-11-08
期刊：
影响因子：
--
通讯作者：
Li J
中科院分区：
文献类型：
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作者：
Qu Y;Wu J;Deng JX;Zhang YP;Liang WY;Jiang ZL;Yu QH;Li J
风湿性关节炎（RA）是一种慢性自身免疫性疾病，可引起关节炎症和破坏，并增加心血管疾病的风险。RA滑膜成纤维细胞（RASFs）参与RA的进展并释放促炎细胞因子。另一方面，microRNA（miRs）可能有助于控制免疫和非免疫细胞的炎症反应。因此，我们的研究使用慢病毒表达载体来测试miR-126过表达对RASF增殖和凋亡的影响。荧光素酶实验验证了miR-126与PIK 3R 2基因的靶向性关系。将抗miR-126和PIK 3R 2 siRNA共转染至RASFs以鉴定PIK 3R 2是否直接参与miR-126诱导的RASFs的增殖和凋亡。采用实时荧光定量PCR（Real-time polymerase chain reaction，PCR）检测miR-126和PIK 3R 2的表达。MTT法检测细胞增殖情况。流式细胞术检测细胞凋亡和细胞周期。Western blotting检测PIK 3R 2、PI 3 K、AKT和p-AKT蛋白表达。Lv-miR-126感染RASFs后，miR-126的相对表达量显著增加。miR-126促进RASF增殖并抑制凋亡。在过表达miR-126的RASF中，PIK 3R 2水平降低，而总PI 3 K和p-AKT水平升高。与单独的抗miR-126处理相比，抗miR-126和PIK 3R 2 siRNA的共转染也增加了PI 3 K和p-AKT水平以及RASF增殖并减少了凋亡。最后，荧光素酶报告基因测定显示miR-126靶向PIK 3R 2。我们的数据表明，miR-126在RASF中的过表达抑制PIK 3R 2的表达，并促进增殖，同时抑制凋亡。这表明抑制miR-126可以在RA的治疗中产生治疗益处。
Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes inflammation and destruction of the joints as well as an increased risk of cardiovascular disease. RA synovial fibroblasts (RASFs) are involved in the progression of RA and release pro-inflammatory cytokines. On the other hand, microRNAs (miRs) may help control the inflammatory response of immune and non-immune cells. Therefore, our study used lentiviral expression vectors to test the effects of miR-126 overexpression on RASF proliferation and apoptosis. Luciferase experiments verified the targeting relationship between miR-126 and PIK3R2 gene. The co-transfection of anti-miR-126 and PIK3R2 siRNA to RASFs were used to identify whether PIK3R2 was directly involved in proliferation and apoptosis of miR-126-induced RASFs. Real-time polymerase chain reaction (PCR) was used to detect miR-126 and PIK3R2 expressions. MTT assay was used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis and cell cycle. Western blotting was used to detect PIK3R2, PI3K, AKT and p-AKT proteins. After Lv-miR-126 infected RASFs, the relative expression of miR-126 was significantly enhanced. MiR-126 promoted RASF proliferation and inhibited apoptosis. Levels of PIK3R2 decreased while total PI3K and p-AKT levels increased in RASFs overexpressing miR-126. Co-transfection of anti-miR-126 and PIK3R2 siRNA also increased PI3K and p-AKT levels as well as RASF proliferation and reduced apoptosis, as compared to anti-miR-126 treatment alone. Finally, luciferase reporter assays showed that miR-126 targeted PIK3R2. Our data indicate that miR-126 overexpression in RASFs inhibits PIK3R2 expression and promotes proliferation while inhibiting apoptosis. This suggests inhibiting miR-126 may yield therapeutic benefits in the treatment of RA.