Mechanistic study on lncRNA XIST/miR-124-3p/ITGB1 axis in renal fibrosis in obstructive nephropathy

IF 3.3 3区生物学 Q3 CELL BIOLOGY

Experimental cell research Pub Date : 2024-08-09 DOI:10.1016/j.yexcr.2024.114194

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引用次数: 0

Abstract

Objective

The purpose of this study was to investigate the role and possible mechanism of lncRNA XIST in renal fibrosis and to provide potential endogenous targets for renal fibrosis in obstructive nephropathy (ON).

Methods

The study included 50 cases of ON with renal fibrosis (samples taken from patients undergoing nephrectomy due to ON) and 50 cases of normal renal tissue (samples taken from patients undergoing total or partial nephrectomy due to accidental injury, congenital malformations, and benign tumors). Treatment of human proximal renal tubular epithelium (HK-2) cells with TGF-β1 simulated renal fibrosis in vitro. Cell viability and proliferation were measured by CCK-8 and EdU, and cell migration was measured by transwell. XIST, miR-124-3p, ITGB1, and epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, α-SMA, and fibronectin) were detected by PCR and immunoblot. The targeting relationship between miR-124-3p and XIST or ITGB1 was verified by starBase and dual luciferase reporter gene experiments. In addition, The left ureter was ligated in mice as a model of unilateral ureteral obstruction (UUO), and the renal histopathology was observed by HE staining and Masson staining.

Results

ON patients with renal fibrosis had elevated XIST and ITGB1 levels and reduced miR-124-3p levels. The administration of TGF-β1 exhibited a dose-dependent promotion of HK-2 cell viability, proliferation, migration, and EMT. Conversely, depleting XIST or enhancing miR-124-3p hindered HK-2 cell viability, proliferation, migration, and EMT in TGF-β1-damaged HK-2 cells HK-2 cells. XIST functioned as a miR-124-3p sponge. Additionally, miR-124-3p negatively regulated ITGB1 expression. Elevating ITGB1 weakened the impact of XIST depletion on TGF-β1-damaged HK-2 cells. Down-regulating XIST improved renal fibrosis in UUO mice.

Conclusion

XIST promotes renal fibrosis in ON by elevating miR-124-3p and reducing ITGB1 expressions.

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lncRNA XIST/miR-124-3p/ITGB1 轴在阻塞性肾病肾纤维化中的作用机制研究

目的本研究旨在探讨lncRNA XIST在肾脏纤维化中的作用和可能机制，并为阻塞性肾病（ON）肾脏纤维化提供潜在的内源性靶点：研究包括50例伴有肾纤维化的阻塞性肾病（样本取自因阻塞性肾病而接受肾切除术的患者）和50例正常肾组织（样本取自因意外伤害、先天性畸形和良性肿瘤而接受全肾或部分肾切除术的患者）。用 TGF-β1 处理人近端肾小管上皮细胞（HK-2），在体外模拟肾脏纤维化。用 CCK-8 和 EdU 测定细胞活力和增殖，用 transwell 测定细胞迁移。通过 PCR 和免疫印迹检测了 XIST、miR-124-3p、ITGB1 和上皮-间质转化（EMT）相关蛋白（E-cadherin、α-SMA 和纤维连接蛋白）。通过starBase和双荧光素酶报告基因实验验证了miR-124-3p与XIST或ITGB1之间的靶向关系。此外，小鼠左输尿管结扎作为单侧输尿管梗阻（UUO）模型，并通过 HE 染色和 Masson 染色观察肾组织病理学：结果：ON肾纤维化患者的XIST和ITGB1水平升高，miR-124-3p水平降低。服用 TGF-β1 对 HK-2 细胞的活力、增殖、迁移和 EMT 有剂量依赖性的促进作用。相反，在受到 TGF-β1 损伤的 HK-2 细胞中，耗尽 XIST 或增强 miR-124-3p 会阻碍 HK-2 细胞的活力、增殖、迁移和 EMT。XIST 起着 miR-124-3p 海绵的作用。此外，miR-124-3p 还能负向调节 ITGB1 的表达。ITGB1 的升高削弱了去掉 XIST 对 TGF-β1 损伤的 HK-2 细胞的影响。下调 XIST 可改善 UUO 小鼠的肾纤维化：结论：XIST通过提高miR-124-3p和降低ITGB1的表达促进ON小鼠的肾脏纤维化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Experimental cell research 医学-细胞生物学

CiteScore

7.20

自引率

0.00%

发文量

295

审稿时长

30 days

期刊介绍： Our scope includes but is not limited to areas such as: Chromosome biology; Chromatin and epigenetics; DNA repair; Gene regulation; Nuclear import-export; RNA processing; Non-coding RNAs; Organelle biology; The cytoskeleton; Intracellular trafficking; Cell-cell and cell-matrix interactions; Cell motility and migration; Cell proliferation; Cellular differentiation; Signal transduction; Programmed cell death.