Cargando…

Analysis of the promoter region of the human miR-32 gene in colorectal cancer

The pathogenesis of colorectal cancer (CRC) is poorly understood. MicroRNA (miR)-32 upregulation in CRC tissues was previously reported, where it increased the proliferation, migration and invasion, and reduced apoptosis of CRC cells by inhibiting the expression of phosphatase and tensin homolog (PT...

Descripción completa

Detalles Bibliográficos
Autores principales: Wu, Weiyun, Tan, Wenkai, Ye, Shicai, Zhou, Yu, Quan, Juanhua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6403515/
https://www.ncbi.nlm.nih.gov/pubmed/30881496
http://dx.doi.org/10.3892/ol.2019.10042
_version_ 1783400623764406272
author Wu, Weiyun
Tan, Wenkai
Ye, Shicai
Zhou, Yu
Quan, Juanhua
author_facet Wu, Weiyun
Tan, Wenkai
Ye, Shicai
Zhou, Yu
Quan, Juanhua
author_sort Wu, Weiyun
collection PubMed
description The pathogenesis of colorectal cancer (CRC) is poorly understood. MicroRNA (miR)-32 upregulation in CRC tissues was previously reported, where it increased the proliferation, migration and invasion, and reduced apoptosis of CRC cells by inhibiting the expression of phosphatase and tensin homolog (PTEN). However, the mechanism underlying miR-32 upregulation remains unknown. miR-32 is an intronic miRNA located within intron 14 of the transmembrane protein 245 gene (TMEM245). The present study aimed to elucidate the biological pathways underlying miR-32 regulation in CRC. A truncated promoter containing the 5′-flanking region of TMEM245/miR-32 gene was constructed. The promoter region was analyzed by dual luciferase reporter assay in CRC cells. DNA pull-down assay and mass spectrometry (MS) were used to identify proteins binding to the core promoter. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and transcription factor (TF) analyses were used to identify the binding proteins. The −320 to −1 bp fragment of the 5′-flanking region exhibited the highest luciferase activity. The regions spanning −606 to −320 bp exhibited a significant decrease in luciferase activity, compared with the −320 to −1 bp fragment. DNA pull-down assay and MS revealed 403 potential miR-32 promoter binding proteins. GO and KEGG pathway analysis indicated that these proteins were involved in numerous physiological and biochemical processes, including ‘structural molecule activity’, ‘RNA binding’, ‘small molecule metabolic process’ and ‘biogenesis’. Furthermore, TF analysis revealed 10 potential interacting TFs, including SMAD family member 1 (SMAD1), signal transducer and activator of transcription 1 (STAT1) and forkhead box K1 (Foxk1). These results suggested that the core promoter region may be located within-320 to −1 bp of the 5′-flanking region of TMEM245/miR-32 gene, while the region from −606 to −320 bp may harbor repressive regulatory elements. The TFs SMAD1, STAT1 and Foxk1 may be involved in the transcriptional regulation of miR-32.
format Online
Article
Text
id pubmed-6403515
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher D.A. Spandidos
record_format MEDLINE/PubMed
spelling pubmed-64035152019-03-15 Analysis of the promoter region of the human miR-32 gene in colorectal cancer Wu, Weiyun Tan, Wenkai Ye, Shicai Zhou, Yu Quan, Juanhua Oncol Lett Articles The pathogenesis of colorectal cancer (CRC) is poorly understood. MicroRNA (miR)-32 upregulation in CRC tissues was previously reported, where it increased the proliferation, migration and invasion, and reduced apoptosis of CRC cells by inhibiting the expression of phosphatase and tensin homolog (PTEN). However, the mechanism underlying miR-32 upregulation remains unknown. miR-32 is an intronic miRNA located within intron 14 of the transmembrane protein 245 gene (TMEM245). The present study aimed to elucidate the biological pathways underlying miR-32 regulation in CRC. A truncated promoter containing the 5′-flanking region of TMEM245/miR-32 gene was constructed. The promoter region was analyzed by dual luciferase reporter assay in CRC cells. DNA pull-down assay and mass spectrometry (MS) were used to identify proteins binding to the core promoter. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and transcription factor (TF) analyses were used to identify the binding proteins. The −320 to −1 bp fragment of the 5′-flanking region exhibited the highest luciferase activity. The regions spanning −606 to −320 bp exhibited a significant decrease in luciferase activity, compared with the −320 to −1 bp fragment. DNA pull-down assay and MS revealed 403 potential miR-32 promoter binding proteins. GO and KEGG pathway analysis indicated that these proteins were involved in numerous physiological and biochemical processes, including ‘structural molecule activity’, ‘RNA binding’, ‘small molecule metabolic process’ and ‘biogenesis’. Furthermore, TF analysis revealed 10 potential interacting TFs, including SMAD family member 1 (SMAD1), signal transducer and activator of transcription 1 (STAT1) and forkhead box K1 (Foxk1). These results suggested that the core promoter region may be located within-320 to −1 bp of the 5′-flanking region of TMEM245/miR-32 gene, while the region from −606 to −320 bp may harbor repressive regulatory elements. The TFs SMAD1, STAT1 and Foxk1 may be involved in the transcriptional regulation of miR-32. D.A. Spandidos 2019-04 2019-02-14 /pmc/articles/PMC6403515/ /pubmed/30881496 http://dx.doi.org/10.3892/ol.2019.10042 Text en Copyright: © Wu et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Wu, Weiyun
Tan, Wenkai
Ye, Shicai
Zhou, Yu
Quan, Juanhua
Analysis of the promoter region of the human miR-32 gene in colorectal cancer
title Analysis of the promoter region of the human miR-32 gene in colorectal cancer
title_full Analysis of the promoter region of the human miR-32 gene in colorectal cancer
title_fullStr Analysis of the promoter region of the human miR-32 gene in colorectal cancer
title_full_unstemmed Analysis of the promoter region of the human miR-32 gene in colorectal cancer
title_short Analysis of the promoter region of the human miR-32 gene in colorectal cancer
title_sort analysis of the promoter region of the human mir-32 gene in colorectal cancer
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6403515/
https://www.ncbi.nlm.nih.gov/pubmed/30881496
http://dx.doi.org/10.3892/ol.2019.10042
work_keys_str_mv AT wuweiyun analysisofthepromoterregionofthehumanmir32geneincolorectalcancer
AT tanwenkai analysisofthepromoterregionofthehumanmir32geneincolorectalcancer
AT yeshicai analysisofthepromoterregionofthehumanmir32geneincolorectalcancer
AT zhouyu analysisofthepromoterregionofthehumanmir32geneincolorectalcancer
AT quanjuanhua analysisofthepromoterregionofthehumanmir32geneincolorectalcancer