Open Access

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

  • Authors:
    • Weiyun Wu
    • Wenkai Tan
    • Shicai Ye
    • Yu Zhou
    • Juanhua Quan
  • View Affiliations

  • Published online on: February 14, 2019     https://doi.org/10.3892/ol.2019.10042
  • Pages: 3743-3750
  • Copyright: © Wu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

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.
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April-2019
Volume 17 Issue 4

Print ISSN: 1792-1074
Online ISSN:1792-1082

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Spandidos Publications style
Wu W, Tan W, Ye S, Zhou Y and Quan J: Analysis of the promoter region of the human miR‑32 gene in colorectal cancer. Oncol Lett 17: 3743-3750, 2019
APA
Wu, W., Tan, W., Ye, S., Zhou, Y., & Quan, J. (2019). Analysis of the promoter region of the human miR‑32 gene in colorectal cancer. Oncology Letters, 17, 3743-3750. https://doi.org/10.3892/ol.2019.10042
MLA
Wu, W., Tan, W., Ye, S., Zhou, Y., Quan, J."Analysis of the promoter region of the human miR‑32 gene in colorectal cancer". Oncology Letters 17.4 (2019): 3743-3750.
Chicago
Wu, W., Tan, W., Ye, S., Zhou, Y., Quan, J."Analysis of the promoter region of the human miR‑32 gene in colorectal cancer". Oncology Letters 17, no. 4 (2019): 3743-3750. https://doi.org/10.3892/ol.2019.10042