Document Type

Article

Journal Title

Journal of cellular biochemistry

Publication Date

8-2010

Volume

110

Abstract

MicroRNAs regulate pathways contributing to oncogenesis, and thus the mechanisms causing dysregulation of microRNA expression in cancer are of significant interest. Mature mir-29b levels are decreased in malignant cells, and this alteration promotes the malignant phenotype, including apoptosis resistance. However, the mechanism responsible for mir-29b suppression is unknown. Here, we examined mir-29 expression from chromosome 7q32 using cholangiocarcinoma cells as a model for mir-29b downregulation. Using 5' rapid amplification of cDNA ends, the transcriptional start site was identified for this microRNA locus. Computational analysis revealed the presence of two putative E-box (Myc-binding) sites, a Gli-binding site, and four NF-kappaB-binding sites in the region flanking the transcriptional start site. Promoter activity in cholangiocarcinoma cells was repressed by transfection with c-Myc, consistent with reports in other cell types. Treatment with the hedgehog inhibitor cyclopamine, which blocks smoothened signaling, increased the activity of the promoter and expression of mature mir-29b. Mutagenesis analysis and gel shift data are consistent with a direct binding of Gli to the mir-29 promoter. Finally, activation of NF-kappaB signaling, via ligation of Toll-like receptors, also repressed mir-29b expression and promoter function. Of note, activation of hedgehog, Toll-like receptor, and c-Myc signaling protected cholangiocytes from TRAIL-induced apoptosis. Thus, in addition to c-Myc, mir-29 expression can be suppressed by hedgehog signaling and inflammatory pathways, both commonly activated in the genesis of human malignancies.

MeSH Headings

Binding Sites, Cell Line, Cell Line, Tumor, Gene Expression Regulation, Hedgehog Proteins, Humans, MicroRNAs, NF-kappa B, Promoter Regions, Genetic, Protein Binding, Proto-Oncogene Proteins c-myc, Signal Transduction, Transcription Factors, Transcription Initiation Site, Transcription, Genetic, Veratrum Alkaloids

ISSN

1097-4644

Comments

This is the pre-peer reviewed version of the following article: Mott JL*, Kurita S, Cazanave SC, Bronk SF, Werneburg NW, Fernandez-Zapico ME. (2010) Transcriptional Suppression of mir-29b-1/mir-29a Promoter by c-Myc, Hedgehog, and NF-kappaB. Journal of Cellular Biochemistry, 110(5):1155-64, which has been published in final form at 10.1002/jcb.22630.

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