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PMID: 19264808 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, N.I.H., Intramural

Estradiol downregulates miR-21 expression and increases miR-21 target gene expression in MCF-7 breast cancer cells.

Nucleic acids research ·Vol. 37 ·No. 8 ·2009-05-00 ·Pages 2584-95

Wickramasinghe NS, Manavalan TT, Dougherty SM, Riggs KA, Li Y, Klinge CM

Abstract

Select changes in microRNA (miRNA) expression correlate with estrogen receptor alpha (ER alpha) expression in breast tumors. miR-21 is higher in ER alpha positive than negative tumors, but no one has examined how estradiol (E(2)) regulates miR-21 in breast cancer cells. Here we report that E(2) inhibits miR-21 expression in MCF-7 human breast cancer cells. The E(2)-induced reduction in miR-21 was inhibited by 4-hydroxytamoxifen (4-OHT), ICI 182 780 (Faslodex), and siRNA ER alpha indicating that the suppression is ER alpha-mediated. ER alpha and ER beta agonists PPT and DPN inhibited and 4-OHT increased miR-21 expression. E(2) increased luciferase activity from reporters containing the miR-21 recognition elements from the 3'-UTRs of miR-21 target genes, corroborating that E(2) represses miR-21 expression resulting in a loss of target gene suppression. The E(2)-mediated decrease in miR-21 correlated with increased protein expression of endogenous miR-21-targets Pdcd4, PTEN and Bcl-2. siRNA knockdown of ER alpha blocked the E(2)-induced increase in Pdcd4, PTEN and Bcl-2. Transfection of MCF-7 cells with antisense (AS) to miR-21 mimicked the E(2)-induced increase in Pdcd4, PTEN and Bcl-2. These results are the first to demonstrate that E(2) represses the expression of an oncogenic miRNA, miR-21, by activating estrogen receptor in MCF-7 cells.

MeSH Terms
3' Untranslated Regions/chemistry Apoptosis Regulatory Proteins/genetics Breast Neoplasms/genetics,metabolism Cell Line, Tumor Cycloheximide/pharmacology Dactinomycin/pharmacology Down-Regulation Estradiol/analogs & derivatives,pharmacology Estrogen Antagonists/pharmacology Estrogen Receptor alpha/agonists,antagonists & inhibitors,genetics Estrogen Receptor beta/agonists,antagonists & inhibitors,genetics Female Fulvestrant Gene Expression Regulation, Neoplastic Genes, Reporter Humans MicroRNAs/antagonists & inhibitors,genetics,metabolism Nucleic Acid Synthesis Inhibitors/pharmacology Promoter Regions, Genetic Protein Synthesis Inhibitors/pharmacology RNA, Antisense/metabolism RNA-Binding Proteins/genetics Tamoxifen/analogs & derivatives,pharmacology ras GTPase-Activating Proteins/genetics
Chemicals
3' Untranslated Regions Apoptosis Regulatory Proteins Estrogen Antagonists Estrogen Receptor alpha Estrogen Receptor beta MIRN21 microRNA, human MicroRNAs Nucleic Acid Synthesis Inhibitors PDCD4 protein, human Protein Synthesis Inhibitors RNA, Antisense RNA-Binding Proteins ras GTPase-Activating Proteins Tamoxifen afimoxifene Dactinomycin Fulvestrant Estradiol Cycloheximide
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wickramasinghe Nalinie S
Department of Biochemistry & Molecular Biology, Center for Genetics and Molecular Medicine, University of Louisville School of Medicine, Louisville, KY 40292, USA.
Manavalan Tissa T
Dougherty Susan M
Riggs Krista A
Li Yong
Klinge Carolyn M
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkp117
Published
2009-05-00
Epub
2009-00-05
Pages
2584-95
Language
English
Region
England
NLM ID
0411011
PMCID
PMC2677875
Subset
IM
Grants
NCI NIH HHS · R21 CA124811 · United States
NIEHS NIH HHS · T32 ES011564 · United States
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1093/nar/gkp117 PMC Full Text View journal details
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