Related Experiment Video
Updated: Jun 17, 2025
MicroRNA In situ Hybridization for Formalin Fixed Kidney Tissues
Published on: November 30, 2013
Mechanistic study on lncRNA XIST/miR-124-3p/ITGB1 axis in renal fibrosis in obstructive nephropathy
ChiTeng Zhang1, KangNing Wang2, Xiang Chen2
1Department of Urology Surgery, The Second Affiliated Hospital, Hengyang Medical School, University of South China, Hengyang City, Hunan Province, 421001, China.
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.
Insights
Long non-coding RNA XIST promotes renal fibrosis by impacting miR-124-3p and ITGB1 levels. Down-regulating XIST shows therapeutic potential for obstructive nephropathy (ON) and renal fibrosis.
Area of Science:
- Nephrology
- Molecular Biology
- Biochemistry
Background:
- Renal fibrosis is a common pathological process in various chronic kidney diseases.
- Obstructive nephropathy (ON) is a significant cause of end-stage renal disease, often leading to renal fibrosis.
- Identifying key molecular mechanisms and therapeutic targets for renal fibrosis is crucial.
Purpose of the Study:
- To investigate the role of long non-coding RNA XIST in renal fibrosis associated with ON.
- To elucidate the underlying molecular mechanism involving XIST, miR-124-3p, and ITGB1.
- To explore XIST as a potential therapeutic target for renal fibrosis.
Main Methods:
- Analysis of XIST, miR-124-3p, and ITGB1 expression in ON patient samples and TGF-β1-induced HK-2 cells.
- Assessment of cell viability, proliferation, migration, and epithelial-mesenchymal transition (EMT) markers.
- Validation of molecular interactions using dual luciferase reporter gene assays and in vivo mouse models of unilateral ureteral obstruction (UUO).
Main Results:
- Elevated XIST and ITGB1, with decreased miR-124-3p, observed in ON patients and TGF-β1-treated cells.
- XIST depletion or miR-124-3p enhancement inhibited TGF-β1-induced HK-2 cell viability, proliferation, migration, and EMT.
- XIST acted as a miR-124-3p sponge, and miR-124-3p negatively regulated ITGB1; ITGB1 elevation counteracted XIST depletion effects.
- Down-regulation of XIST ameliorated renal fibrosis in UUO mice.
Conclusions:
- XIST promotes renal fibrosis in ON by sponging miR-124-3p and subsequently increasing ITGB1 expression.
- Targeting XIST presents a potential therapeutic strategy for mitigating renal fibrosis in ON.
Related Concept Videos
02:39lncRNA - Long Non-coding RNAs
03:17Inheritance of Chromatin Structures