microRNA-126 targeting PIK3R2 promotes rheumatoid arthritis synovial fibro-blasts proliferation and resistance to apoptosis by regulating PI3K/AKT pathway
microRNA-126 targeting PIK3R2 promotes rheumatoid arthritis synovial fibro-blasts proliferation and resistance to apoptosis by regulating PI3K/AKT pathway
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DOI：
10.1016/j.yexmp.2015.12.015
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发表时间：
2016-02-01
影响因子：
3.6
通讯作者：
Zhao, Dong-Bao
中科院分区：
文献类型：
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作者：
Gao, Jie;Zhou, Xiao-Li;Zhao, Dong-Bao
目的：本研究旨在探讨靶向PIK 3R 2基因的microRNA-126（miR-126）通过调控PI 3 K/AKT信号通路对类风湿关节炎滑膜成纤维细胞（RASFs）增殖和凋亡的影响。通过TargetScan收集miR-126的靶基因，通过双荧光素酶报告基因分析系统确定PIIC 3R 2为miR-126的直接靶基因。实验分为空白对照组、miR-126模拟物组、miR-126模拟物对照组、miR-126抑制剂组和miR-126抑制剂对照组。结果：与健康人相比，RA患者滑膜组织中miR-126 mRNA表达水平升高，PIK 3R 2 mRNA表达水平降低。Pearson相关分析显示miR-126与PIK 3R 2 mRNA表达呈负相关（均P <0. 05）。与空白组相比，miR-126模拟物组S期和G2/M期细胞比例增加，细胞凋亡率降低; miR-126抑制剂组G 0/G1期和G2/M期细胞比例增加，细胞凋亡率升高（均P <0. 05）。与空白对照组相比，miR-126模拟物组PIK 3R 2蛋白表达降低，PI 3 K和p-AKT蛋白表达升高miR-126抑制剂组PIK 3R 2蛋白表达增加，PI 3 K和p-AKT蛋白表达减少，（均P < 0.05）。结论：我们的研究表明，下调miR-126可能通过靶向PIK 3R 2间接抑制PI 3 K/AKT信号通路，从而破坏RASFs生长和死亡之间的失衡，这可能在临床上有助于发现针对RA患者的miR-126功能的治疗策略。(C)2015 Elsevier Inc. All rights reserved.
Objective: The purpose of our study was to elucidate the impact of microRNA-126 (miR-126) targeting PIK3R2 gene on cell proliferation and apoptosis of rheumatoid arthritis synovial fibro-blasts (RASFs) by regulating PI3K/AKT signal pathway.Methods: The synovial tissue samples of this study were from 55 RA patients undergoing joint replacement and 27 healthy people undergoing joint repair due to trauma. The target genes of miR-126 were collected by the TargetScan and PIIC3R2 as the direct target gene of miR-126 was confirmed by dual-luciferase reporter assay system. Our experiment had five groups including the blank control, miR-126 mimic, miR-126 mimic control, miR-126 inhibitor and miR-126 inhibitor control groups. Additionally, real-time quantitative polymerase chain reaction (RT-qPCR), Western-Blot, cell counting kit (CCK-8) and flow cytometry were carried out in this study.Results: Compared with healthy individuals, the RA patients had increased miR-126, but decreased PIK3R2 mRNA expressions in the synovial tissues. Pearson correlation analysis indicated that miR-126 expression was negatively correlated with PIK3R2 mRNA expression (all P < 0.05). When compared with the blank group respectively, the miR-126 mimic group had raising cell proportions in S and G2/M phases with reduced rate of cell apoptosis, while the miR-126 inhibitor group had raising cell proportions in G0/G1 and G2/M phases with increased rate of cell apoptosis (all P < 0.05). Besides, compared with the blank control group, the miR-126 mimic group had declined expression of PIK3R2 protein with ascended expression of PI3K and p-AKT (all P < 0.05), while the miR-126 inhibitor group had increased expression of PIK3R2 protein with decreased expression of PI3K and p-AKT (all P < 0.05).Conclusion: Our study demonstrated that down-regulation of miR-126 may indirectly inhibit PI3K/AKT signaling pathway to disrupt the imbalance between growth and death of RASFs by targeting PIK3R2, which may be clinically helpful to find therapeutic strategies directed toward miR-126 function for RA patients. (C) 2015 Elsevier Inc. All rights reserved.