Abstract
The Wnt/beta-catenin/TCF4 pathway plays critical roles in the maintenance of small intestinal epithelium; however, downstream targets of the beta-catenin/TCF4 complex are not extensively characterized. We identified miR-30e as an immediate target activated by the beta-catenin/TCF4 complex. miR-30e was detected in the peri-nuclear region of the intestinal crypt IEC-6 cells. Bioinformatics analysis revealed clustered beta-catenin/TCF4 binding sites within the miR-30e promoter region. This promoter region was cloned into pGL3-control luciferase reporter vector, with the enhancer region removed. Transfection of pCMV-SPORT6-beta-catenin expression vector dose-dependently increased luciferase activity, and co-transfection of pCMV-SPORT6-TCF4 expression vector further enhanced the promoter activity. Dexamethasone-induced IEC-6 cells differentiation caused a 2.5-fold increase in miR-30e expression, and upon beta-catenin siRNA transfection, miR-30e increased 1.3-fold. Electrophoretic mobility shift assay and chromatin immunoprecipitation assay confirmed the binding between beta-catenin/TCF4 complexes from IEC-6 nuclear extracts and the putative sequences in the miR-30e promoter. These results demonstrate that beta-catenin/TCF4 transactivates miR-30e during intestinal cell differentiation.
Original language | English (US) |
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Pages (from-to) | 2969-2978 |
Number of pages | 10 |
Journal | Cellular and Molecular Life Sciences |
Volume | 67 |
Issue number | 17 |
DOIs | |
State | Published - Sep 2010 |
Keywords
- Beta-catenin
- Differentiation
- IEC-6 cells
- Intestine
- MiR-30e
- TCF4
ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
- Molecular Medicine
- Pharmacology
- Cellular and Molecular Neuroscience
- Medicine(all)