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MicroRNA-193a-3p participates in the progression of rheumatoid arthritis by regulating proliferation and apoptosis of MH7A cells through targeting IGFBP5.

基因敲除 小RNA 细胞凋亡 流式细胞术 荧光素酶 下调和上调 报告基因 肿瘤坏死因子α 化学 细胞生长 转染 分子生物学 癌症研究 基因表达 免疫学 基因 生物 生物化学
作者
Qu Sp,Li Gw,Hao Ma,Qi Xing
出处
期刊:PubMed 卷期号:23 (11): 4850-4857 被引量:11
标识
DOI:10.26355/eurrev_201906_18072
摘要

This study aims to explore the regulatory effect of microRNA-193a-3p on rheumatoid arthritis (RA) and its underlying mechanism.Expression level of microRNA-193a-3p in synovial tissues extracted from 30 RA patients and healthy controls was detected by quantitative Real-time polymerase chain reaction (qRT-PCR). MH7A cells were subjected to TNF-α induction for constructing the in vitro RA model. After transfection of microRNA-193a-3p inhibitor in MH7A cells, proliferation and apoptosis were detected by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively. Enzyme-linked immunosorbent assay (ELISA) was conducted to determine levels of interleukin 6 (IL-6) and IL-8 in MH7A cells. Subsequently, the dual-luciferase reporter gene assay was carried out to verify the binding condition between microRNA-193a-3p and IGFBP5. Rescue experiments were conducted to evaluate the proliferation and apoptosis of MH7A cells with knockdown of microRNA-193a-3p and IGFBP5.MicroRNA-193a-3p was highly expressed in synovial tissues of RA patients and TNF-α-induced MH7A cells than those of controls. TNF-α induction significantly increased the proliferative rate of MH7A cells, reaching the peak at 96 h. After knockdown of microRNA-193a-3p, the promoted proliferation by TNF-α induction was significantly inhibited. In addition, TNF-α induction significantly inhibited the apoptosis of MH7A cells. After inhibition of microRNA-193a-3p expression, the inhibited apoptosis by TNF-α induction remarkably increased. TNF-α induction upregulated levels of IL-6 and IL-8 in MH7A cells, which were remarkably reduced after the microRNA-193a-3p knockdown. Dual-luciferase reporter gene assay confirmed that IGFBP5 could bind to microRNA-193a-3p, and its expression was negatively regulated by microRNA-193a-3p. The regulatory effects of microRNA-193a-3p on proliferation and apoptosis of MH7A cells were reversed by IGFBP5 knockdown.MicroRNA-193a-3p is highly expressed in the synovial tissues and cells of rheumatoid arthritis. MicroRNA-193a-3p participates in the process of rheumatoid arthritis by regulating the proliferation, apoptosis and inflammatory response of MH7A cells through targeting IGFBP5.
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