Circ_FURIN敲低通过海绵miR-423-5p降低多囊卵巢综合征中MTM1的表达来缓解睾酮诱导的人卵巢颗粒样肿瘤细胞紊乱。

Xia Xu, Rui Guan, Ke Gong, Huaibing Xie, Lei Shi
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引用次数: 7

摘要

背景:多囊卵巢综合征(PCOS)是育龄妇女中一种常见的内分泌疾病。环状RNA (circRNA)驱动PCOS发展的机制尚不清楚。因此,本研究旨在探索一种新型circRNA circ_FURIN在PCOS细胞模型中的作用及其潜在机制。方法:用睾酮(TTR)治疗人卵巢颗粒样肿瘤细胞(KGN),建立PCOS细胞模型。采用实时定量聚合酶链式反应(qRT-PCR)检测circ_FURIN、microRNA-423-5p (miR-423-5p)和肌管蛋白1 (MTM1)的RNA表达。Western blot检测蛋白表达。采用5-乙基-29脱氧尿苷法、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑(MTT)法和流式细胞术检测细胞周期。采用流式细胞术定量检测细胞凋亡。miR-423-5p与circ_FURIN或MTM1之间的相互作用通过双荧光素酶报告基因和RNA下拉试验进行鉴定。结果:与对照组相比,PCOS患者卵巢皮质组织和经trr处理的KGN细胞中,Circ_FURIN和MTM1表达显著上调,而miR-423-5p表达下调。Circ_FURIN缺失减轻了trr诱导的增殖抑制和细胞凋亡促进。此外,敲低circ_FURIN的靶miRNA miR-423-5p,可在TTR治疗下恢复circ_FURIN敲低介导的效应。MiR-423-5p通过结合MTM1缓解trr诱导的细胞紊乱。此外,circ_FURIN通过miR-423-5p调节MTM1的表达。结论:Circ_FURIN沉默可通过miR-423-5p/MTM1通路保护人卵巢颗粒样肿瘤细胞免受ttr诱导的功能障碍。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Circ_FURIN knockdown assuages Testosterone-induced human ovarian granulosa-like tumor cell disorders by sponging miR-423-5p to reduce MTM1 expression in polycystic ovary syndrome.

Circ_FURIN knockdown assuages Testosterone-induced human ovarian granulosa-like tumor cell disorders by sponging miR-423-5p to reduce MTM1 expression in polycystic ovary syndrome.

Circ_FURIN knockdown assuages Testosterone-induced human ovarian granulosa-like tumor cell disorders by sponging miR-423-5p to reduce MTM1 expression in polycystic ovary syndrome.

Circ_FURIN knockdown assuages Testosterone-induced human ovarian granulosa-like tumor cell disorders by sponging miR-423-5p to reduce MTM1 expression in polycystic ovary syndrome.

Background: Polycystic ovary syndrome (PCOS) is a common endocrine disorder among reproductive-age women. The mechanism by which circular RNA (circRNA) drives PCOS development remains unclear. Thus, the study is designed to explore the role of a novel circRNA, circ_FURIN, in the PCOS cell model and the underlying mechanism.

Methods: PCOS cell model was established by treating human ovarian granulosa-like tumor cells (KGN) with Testosterone (TTR). RNA expressions of circ_FURIN, microRNA-423-5p (miR-423-5p) and myotubularin 1 (MTM1) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was checked by Western blot. Cell proliferation was investigated by a 5-Ethynyl-29-deoxyuridine assay, 3-(4,5-Dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis for cell cycle. Apoptotic cells were quantified by flow cytometry analysis for cell apoptosis. The interplay between miR-423-5p and circ_FURIN or MTM1 was identified by dual-luciferase reporter and RNA pull-down assays.

Results: Circ_FURIN and MTM1 expressions were significantly upregulated, whereas miR-423-5p was downregulated in the ovarian cortex tissues of PCOS patients and TTR-treated KGN cells compared with controls. Circ_FURIN depletion relieved TTR-induced proliferation inhibition and apoptosis promotion. Besides, knockdown of miR-423-5p, a target miRNA of circ_FURIN, rescued circ_FURIN knockdown-mediated effects under TTR treatment. MiR-423-5p remitted TTR-induced cell disorders by binding to MTM1. Moreover, circ_FURIN modulated MTM1 expression through miR-423-5p.

Conclusion: Circ_FURIN silencing protected against TTR-induced dysfunction by the miR-423-5p/MTM1 pathway in human ovarian granulosa-like tumor cells.

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