MicroRNA-124 Regulates STAT3 Expression and Is Down-regulated in Colon Tissues of Pediatric Patients With Ulcerative Colitis

• Published in: Gastroenterology (New York, N.Y. 1943) Vol. 145; no. 4; pp. 842 - 852.e2
• Main Authors: Koukos, Georgios, Polytarchou, Christos, Kaplan, Jess L, Morley–Fletcher, Alessio, Gras–Miralles, Beatriz, Kokkotou, Efi, Baril–Dore, Mariah, Pothoulakis, Charalabos, Winter, Harland S, Iliopoulos, Dimitrios
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.2013
• Subjects: 3' Untranslated Regions; Adolescent; Animals; BCLXL; Cell Line, Tumor; Child; Child, Preschool; Colitis, Ulcerative - metabolism; Colon - metabolism; DNA Methylation; Down-Regulation; Gastroenterology and Hepatology; Gene Expression Regulation; Gene Regulation; High-Throughput Screening Assays; Humans; IL6; Infant; Infant, Newborn; Let-7; Male; Mice; Mice, Inbred C57BL; MicroRNAs - physiology; Promoter Regions, Genetic; RNA, Messenger - analysis; STAT3 Transcription Factor - genetics; IBD; mRNA; tyrosine 705; dextran sulfate sodium salt; UC; Gene Regulation; 5-AZA; messenger RNA; ulcerative colitis; Tyr705; interleukin; matrix metallopeptidase 9; BCLXL; polymerase chain reaction; knockout; CD; IL; KO; DSS; VEGF; STAT3; Crohn's disease; signal transducer and activator of transcription 3; MMP9; inflammatory bowel disease; IL6; phosphorylated STAT3; vascular endothelial growth factor; enzyme-linked immunosorbent assay; miR; microRNA; Let-7; PCR; p-STAT3; 5-aza-2′-deoxycytidine; ELISA
• ISSN: 0016-5085; 1528-0012
• DOI: 10.1053/j.gastro.2013.07.001

Background & Aims Altered levels and functions of microRNAs (miRs) have been associated with inflammatory bowel diseases (IBDs), although little is known about their roles in pediatric IBD. We investigated whether colonic mucosal miRs are altered in children with ulcerative colitis (UC). Methods We used a library of 316 miRs to identify those that regulate phosphorylation of signal transducer and activator of transcription 3 (STAT3) in NCM460 human colonocytes incubated with interleukin-6. Levels of miR-124 were measured by real-time polymerase chain reaction analysis of colon biopsies from pediatric and adult patients with UC and patients without IBD (controls), and of HCT-116 colonocytes incubated with 5-aza-2′-deoxycytidine (5-AZA). Methylation of the MIR124 promoter was measured by quantitative methylation-specific polymerase chain reaction. Results Levels of phosphorylated STAT3 and the genes it regulates (encoding vascular endothelial growth factor (VEGF), BCL2, BCLXL, and matrix metallopeptidase 9 [MMP9]) were increased in pediatric patients with UC compared with control tissues. Overexpression of miR-124, let-7, miR-125, miR-26, or miR-101 reduced STAT3 phosphorylation by ≥75% in NCM460 cells; miR-124 had the greatest effect. miR-124 was down-regulated specifically in colon tissues from pediatric patients with UC and directly targeted STAT3 messenger RNA (mRNA). Levels of miR-124 were decreased, whereas levels of STAT3 phosphorylation increased in colon tissues from pediatric patients with active UC compared with those with inactive disease. In addition, levels of miR-124 and STAT3 were inversely correlated in mice with experimental colitis. Down-regulation of miR-124 in tissues from children with UC was attributed to hypermethylation of its promoter region. Incubation of HCT-116 colonocytes with 5-AZA up-regulated miR-124 and reduced levels of STAT3 mRNA. Conclusions miR-124 appears to regulate the expression of STAT3. Reduced levels of miR-124 in colon tissues of children with active UC appear to increase expression and activity of STAT3, which could promote inflammation and the pathogenesis of UC in children.