Cargando…

Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells

The miR-200b~200a~429 gene cluster is a key regulator of EMT and cancer metastasis, however the transcription-based mechanisms controlling its expression during this process are not well understood. We have analyzed the miR-200b~200a~429 locus for epigenetic modifications in breast epithelial and me...

Descripción completa

Detalles Bibliográficos
Autores principales: Attema, Joanne L., Bert, Andrew G., Lim, Yat-Yuen, Kolesnikoff, Natasha, Lawrence, David M., Pillman, Katherine A., Smith, Eric, Drew, Paul A., Khew-Goodall, Yeesim, Shannon, Frances, Goodall, Gregory J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3783398/
https://www.ncbi.nlm.nih.gov/pubmed/24086551
http://dx.doi.org/10.1371/journal.pone.0075517
_version_ 1782285655827546112
author Attema, Joanne L.
Bert, Andrew G.
Lim, Yat-Yuen
Kolesnikoff, Natasha
Lawrence, David M.
Pillman, Katherine A.
Smith, Eric
Drew, Paul A.
Khew-Goodall, Yeesim
Shannon, Frances
Goodall, Gregory J.
author_facet Attema, Joanne L.
Bert, Andrew G.
Lim, Yat-Yuen
Kolesnikoff, Natasha
Lawrence, David M.
Pillman, Katherine A.
Smith, Eric
Drew, Paul A.
Khew-Goodall, Yeesim
Shannon, Frances
Goodall, Gregory J.
author_sort Attema, Joanne L.
collection PubMed
description The miR-200b~200a~429 gene cluster is a key regulator of EMT and cancer metastasis, however the transcription-based mechanisms controlling its expression during this process are not well understood. We have analyzed the miR-200b~200a~429 locus for epigenetic modifications in breast epithelial and mesenchymal cell lines using chromatin immunoprecipitation assays and DNA methylation analysis. We discovered a novel enhancer located approximately 5.1kb upstream of the miR-200b~200a~429 transcriptional start site. This region was associated with the active enhancer chromatin signature comprising H3K4me1, H3K27ac, RNA polymerase II and CpG dinucleotide hypomethylation. Luciferase reporter assays revealed the upstream enhancer stimulated the transcription of the miR-200b~200a~429 minimal promoter region approximately 27-fold in breast epithelial cells. Furthermore, we found that a region of the enhancer was transcribed, producing a short, GC-rich, mainly nuclear, non-polyadenylated RNA transcript designated miR-200b eRNA. Over-expression of miR-200b eRNA had little effect on miR-200b~200a~429 promoter activity and its production did not correlate with miR-200b~200a~429 gene expression. While additional investigations of miR-200b eRNA function will be necessary, it is possible that miR-200b eRNA may be involved in the regulation of miR-200b~200a~429 gene expression and silencing. Taken together, these findings reveal the presence of a novel enhancer, which contributes to miR-200b~200a~429 transcriptional regulation in epithelial cells.
format Online
Article
Text
id pubmed-3783398
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-37833982013-10-01 Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells Attema, Joanne L. Bert, Andrew G. Lim, Yat-Yuen Kolesnikoff, Natasha Lawrence, David M. Pillman, Katherine A. Smith, Eric Drew, Paul A. Khew-Goodall, Yeesim Shannon, Frances Goodall, Gregory J. PLoS One Research Article The miR-200b~200a~429 gene cluster is a key regulator of EMT and cancer metastasis, however the transcription-based mechanisms controlling its expression during this process are not well understood. We have analyzed the miR-200b~200a~429 locus for epigenetic modifications in breast epithelial and mesenchymal cell lines using chromatin immunoprecipitation assays and DNA methylation analysis. We discovered a novel enhancer located approximately 5.1kb upstream of the miR-200b~200a~429 transcriptional start site. This region was associated with the active enhancer chromatin signature comprising H3K4me1, H3K27ac, RNA polymerase II and CpG dinucleotide hypomethylation. Luciferase reporter assays revealed the upstream enhancer stimulated the transcription of the miR-200b~200a~429 minimal promoter region approximately 27-fold in breast epithelial cells. Furthermore, we found that a region of the enhancer was transcribed, producing a short, GC-rich, mainly nuclear, non-polyadenylated RNA transcript designated miR-200b eRNA. Over-expression of miR-200b eRNA had little effect on miR-200b~200a~429 promoter activity and its production did not correlate with miR-200b~200a~429 gene expression. While additional investigations of miR-200b eRNA function will be necessary, it is possible that miR-200b eRNA may be involved in the regulation of miR-200b~200a~429 gene expression and silencing. Taken together, these findings reveal the presence of a novel enhancer, which contributes to miR-200b~200a~429 transcriptional regulation in epithelial cells. Public Library of Science 2013-09-25 /pmc/articles/PMC3783398/ /pubmed/24086551 http://dx.doi.org/10.1371/journal.pone.0075517 Text en © 2013 Attema et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Attema, Joanne L.
Bert, Andrew G.
Lim, Yat-Yuen
Kolesnikoff, Natasha
Lawrence, David M.
Pillman, Katherine A.
Smith, Eric
Drew, Paul A.
Khew-Goodall, Yeesim
Shannon, Frances
Goodall, Gregory J.
Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells
title Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells
title_full Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells
title_fullStr Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells
title_full_unstemmed Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells
title_short Identification of an Enhancer That Increases miR-200b~200a~429 Gene Expression in Breast Cancer Cells
title_sort identification of an enhancer that increases mir-200b~200a~429 gene expression in breast cancer cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3783398/
https://www.ncbi.nlm.nih.gov/pubmed/24086551
http://dx.doi.org/10.1371/journal.pone.0075517
work_keys_str_mv AT attemajoannel identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT bertandrewg identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT limyatyuen identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT kolesnikoffnatasha identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT lawrencedavidm identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT pillmankatherinea identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT smitheric identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT drewpaula identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT khewgoodallyeesim identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT shannonfrances identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells
AT goodallgregoryj identificationofanenhancerthatincreasesmir200b200a429geneexpressioninbreastcancercells