title: Microarray analysis of altered gene expression in ERbeta-overexpressing HEK293 cells.
abstract: Estrogen receptors (ERs), ERalpha and ERbeta, mediate estrogen actions in a broad range of target tissues. With the introduction of microarray techniques, a significant understanding has been gained regarding the interplay between the ERalpha and ERbeta in breast cancer cell lines. To gain a more comprehensive understanding of ERbeta-dependent gene regulation independent of ERalpha, we performed microarray analysis on HEK293/mock and HEK293/ERbeta cells. A total of 332 genes was identified as ERbeta-upregulated genes and 210 identified as ERbeta-downregulated genes. ERbeta-induced and ERbeta-repressed genes were involved in cell-cell signaling, morphogenesis, and cell proliferation. The ERbeta repressive effect on genes related to proliferation was further studied by proliferation assays, where ERbeta expression resulted in a significant decrease in cell proliferation. To identify primary ERbeta target genes, we examined a number of ERbeta-regulated genes using chromatin immunoprecipitation assays for regions bound by ERbeta. Our results showed that ERbeta recruitment was significant to regions associated with 12 genes (IL1RAP, TMSB4X, COLEC12, ENPP2, KLRC1, RERG, RGS16, TNNT2, CYR61, FER1L3, FAM108A1, and CYP4X1), suggesting that these genes are likely to be ERbeta primary target genes. This study has provided novel information on the gene regulatory function of ERbeta independent of ERalpha and identified a number of ERbeta primary target genes. The results of Gene Ontology analysis and proliferation assays are consistent with an antiproliferative role of ERbeta independent of ERalpha.
authors.affiliation: Department of Biosciences and Nutrition, NOVUM, Karolinska Institutet, 141 57, Huddinge, Sweden. chunyan.zhao@ki.se
authors.forename: Chunyan
authors.initials: C
authors.lastname: Zhao
authors.forename: Milica
authors.initials: M
authors.lastname: Putnik
authors.forename: Jan-Ake
authors.initials: JA
authors.lastname: Gustafsson
authors.forename: Karin
authors.initials: K
authors.lastname: Dahlman-Wright
mesh_terms: D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation; D047629:Estrogen Receptor beta / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D014162:Transfection
chemical_list: D047629:Estrogen Receptor beta
references.citation: Proc Natl Acad Sci U S A. 2004 Feb 10;101(6):1566-71
references.pmid: 14745018
references.citation: Nucleic Acids Res. 2004 Jan 1;32(Database issue):D258-61
references.pmid: 14681407
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2604-9
references.pmid: 18272478
references.citation: Nucleic Acids Res. 2001 Jul 15;29(14):2905-19
references.pmid: 11452016
references.citation: PLoS Genet. 2007 Jun;3(6):e87
references.pmid: 17542648
references.citation: Crit Rev Eukaryot Gene Expr. 2002;12(4):237-57
references.pmid: 12641394
references.citation: Carcinogenesis. 2004 Nov;25(11):2067-73
references.pmid: 15205361
references.citation: Biochem Biophys Res Commun. 2001 Jul 13;285(2):340-7
references.pmid: 11444848
references.citation: Genome Biol. 2003;4(10):R70
references.pmid: 14519205
references.citation: Endocrinology. 1998 Oct;139(10):4252-63
references.pmid: 9751507
references.citation: Mol Endocrinol. 2008 Feb;22(2):361-79
references.pmid: 17962383
references.citation: Cancer Res. 2007 Apr 15;67(8):3955-62
references.pmid: 17440111
references.citation: Mol Endocrinol. 2006 Mar;20(3):534-43
references.pmid: 16293641
references.citation: Mol Pharmacol. 1998 Jul;54(1):105-12
references.pmid: 9658195
references.citation: Endocrinology. 2006 Oct;147(10):4831-42
references.pmid: 16809442
references.citation: J Biol Chem. 2001 Apr 27;276(17):14187-94
references.pmid: 11297518
references.citation: Science. 1995 Dec 1;270(5241):1491-4
references.pmid: 7491495
references.citation: Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W249-52
references.pmid: 15215389
references.citation: J Cell Biochem. 2003 Oct 1;90(2):315-26
references.pmid: 14505348
references.citation: J Steroid Biochem Mol Biol. 2000 Nov 30;74(5):311-7
references.pmid: 11162939
references.citation: Mol Cell Endocrinol. 2007 Aug 15;274(1-2):19-29
references.pmid: 17597290
references.citation: Breast Cancer Res. 2007;9(2):R25
references.pmid: 17428314
references.citation: J Biol Chem. 2000 Jun 16;275(24):18447-53
references.pmid: 10749889
references.citation: Nat Genet. 2006 Nov;38(11):1289-97
references.pmid: 17013392
references.citation: Nat Genet. 2000 May;25(1):25-9
references.pmid: 10802651
references.citation: J Biol Chem. 2001 Nov 9;276(45):42259-67
references.pmid: 11533059
references.citation: Cell. 1989 Feb 10;56(3):335-44
references.pmid: 2644044
references.citation: Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):13162-7
references.pmid: 16938840
references.citation: J Biol Chem. 2000 Feb 25;275(8):5379-87
references.pmid: 10681512
references.citation: J Cell Biochem. 2007 Aug 1;101(5):1125-47
references.pmid: 17520659
references.citation: Oncogene. 2003 Aug 7;22(32):5011-20
references.pmid: 12902984
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1339-44
references.pmid: 15677324
references.citation: Oncogene. 2008 Feb 7;27(7):1019-32
references.pmid: 17700529
references.citation: Mol Cell Biol. 2003 Aug;23(16):5867-81
references.pmid: 12897156