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Perturbations of Circulating miRNAs in Irritable Bowel Syndrome Detected Using a Multiplexed High-throughput Gene Expression Platform
Published on: November 30, 2016
The diagnostic value of miR-340-5p in pediatric ulcerative colitis and its molecular mechanism by targeting MAP3K2 to
Fanting Meng1, Xianlong Han2, Lingxia Ge3,4
1Department of Gastroenterology, Children's Hospital of Nanjing Medical University, Nanjing City, Jiangsu Province, 210000, China.
Insights
MicroRNA-340-5p is a potential diagnostic biomarker for pediatric ulcerative colitis (UC). It targets MAP3K2 to regulate inflammation, apoptosis, and oxidative stress in pediatric UC.
Area of Science:
- Gastroenterology
- Molecular Biology
- Biomarker Discovery
Background:
- Pediatric ulcerative colitis (UC) incidence is rising, necessitating novel diagnostic biomarkers and understanding of molecular mechanisms.
- Current research lacks specific biomarkers for early diagnosis and effective treatment of pediatric UC.
Purpose of the Study:
- To investigate the diagnostic value of microRNA-340-5p (miR-340-5p) in pediatric UC.
- To elucidate the molecular mechanism of miR-340-5p in pediatric UC by identifying its target, MAP3K2.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to measure miR-340-5p and MAP3K2 expression in 85 pediatric UC patients and 50 healthy controls.
- Dual-luciferase assay to confirm the interaction between miR-340-5p and MAP3K2.
- In vitro cell models (HT-29 cells) treated with dextran sulfate sodium (DSS) to assess cellular functions via flow cytometry, ELISA, and oxidative stress assays.
Main Results:
- miR-340-5p was significantly downregulated in pediatric UC patients and negatively correlated with disease activity indices (PUCAI, CRP, ESR).
- ROC curve analysis showed an area of 0.908 for miR-340-5p as a diagnostic marker.
- miR-340-5p directly targets MAP3K2, suppressing its expression and mitigating DSS-induced apoptosis, inflammation (reduced TNF-α, IL-6, IL-17, IL-1β), and oxidative stress (reduced MDA, increased GSH).
Conclusions:
- miR-340-5p demonstrates potential as a diagnostic biomarker for pediatric UC.
- The miR-340-5p/MAP3K2 axis plays a crucial role in regulating apoptosis, inflammatory responses, and oxidative stress in pediatric UC.
- Targeting the miR-340-5p/MAP3K2 pathway may offer therapeutic strategies for pediatric UC.
Abstract:
BACKGROUND: The incidence of pediatric ulcerative colitis (UC) is increasing yearly, and it is urgent to explore precise diagnostic biomarkers and molecular mechanisms. This study aims to investigate the diagnostic value of miR-340-5p in pediatric UC and its molecular mechanism mediated through targeting MAP3K2. METHODS: Eighty-five pediatric UC patients and 50 healthy controls were enrolled. The expression levels of miR-340-5p and MAP3K2 were detected by qRT-PCR, and Pearson’s correlation analysis was conducted. An in vitro model was established by inducing HT-29 cells with dextran sulfate sodium. The target was verified by dual-luciferase assay. Flow cytometry, ELISA, and oxidative stress detection were used to verify the cellular functions regulated by the miR-340-5p/MAP3K2 axis. RESULTS: In pediatric patients with UC, miR-340-5p was significantly downregulated and negatively correlated with PUCAI, CRP, and ESR (P < 0.0001). Additionally, the diagnostic area of miR-340-5p under the ROC curve was 0.908. Mechanistically, miR-340-5p directly interacts with MAP3K2, leading to suppressed expression of its mRNA and protein. Functional experiments revealed that miR-340-5p overexpression reversed DSS-induced exacerbated cell apoptosis, reduced levels of TNF-α, IL-6, IL-17, IL-1β, and MDA, and increased GSH. Conversely, the beneficial effects of miR-340-5p were attenuated by oe-MAP3K2 overexpression. CONCLUSIONS: miR-340-5p may serve as a diagnostic biomarker for pediatric UC and exerts its effects by targeting MAP3K2 to regulate cell apoptosis, inflammatory responses, and oxidative stress.
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