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F Wu, Biochemistry and Biophysics, Texas A&M University, College Station, United States
I Ivanov, Veterinary Physiology and Pharmacology, Texas A&M University, College Station, United States
R Xu, Biochemistry and Biophysics, Texas A&M University, College Station, United States
S Safe, Veterinary Physiology & Pharmacology, Texas A&M University, College Station, 77843-4466, United States

Correspondence: Stephen Safe, Email: ssafe{at}cvm.tamu.edu

Abstract

17β-Estradiol (E2) binds estrogen receptor (ER) in MCF-7 cells and increases cell proliferation and survival through induction or repression of multiple genes. ER interactions with DNA-bound specificity protein (Sp) transcription factors is a non-classical genomic estrogenic pathway and the role of Sp transcription factors in mediating hormone-dependent activation or repression of genes in MCF-7 cells was investigated by microarrays and RNA interference. MCF-7 cells were transfected with a non-specific oligonucleotide or a cocktail of small inhibitory RNAs (iSp) which knockdown Sp1, Sp3 and Sp4 proteins and treated with DMSO or 10 nM E2 for 6 hr. E2 induced 62 and repressed 134 genes and the induction or repression was reversed in approximately 62% of the genes in cells transfected with iSp (ER/Sp-dependent), whereas hormonal activation or repression of the remaining genes was unaffected by iSp (Sp-independent). Analysis of the ER/Sp-dependent and Sp-independent genes showed minimal overlap with respect to the GO terms (functional processes) in genes induced or repressed, suggesting that the different genomic pathways may contribute independently to the hormone-induced phenotype in MCF-7 cells.

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