Microarray analysis of altered gene expression in ERbeta-overexpressing HEK293 cells.
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.
Zhao
Chunyan
C
Department of Biosciences and Nutrition, NOVUM, Karolinska Institutet, 141 57, Huddinge, Sweden. chunyan.zhao@ki.se
Putnik
Milica
M
Gustafsson
Jan-Ake
JA
Dahlman-Wright
Karin
K
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
D047629:Estrogen Receptor beta
Proc Natl Acad Sci U S A. 2004 Feb 10;101(6):1566-71
14745018
Nucleic Acids Res. 2004 Jan 1;32(Database issue):D258-61
14681407
Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2604-9
18272478
Nucleic Acids Res. 2001 Jul 15;29(14):2905-19
11452016
PLoS Genet. 2007 Jun;3(6):e87
17542648
Crit Rev Eukaryot Gene Expr. 2002;12(4):237-57
12641394
Carcinogenesis. 2004 Nov;25(11):2067-73
15205361
Biochem Biophys Res Commun. 2001 Jul 13;285(2):340-7
11444848
Genome Biol. 2003;4(10):R70
14519205
Endocrinology. 1998 Oct;139(10):4252-63
9751507
Mol Endocrinol. 2008 Feb;22(2):361-79
17962383
Cancer Res. 2007 Apr 15;67(8):3955-62
17440111
Mol Endocrinol. 2006 Mar;20(3):534-43
16293641
Mol Pharmacol. 1998 Jul;54(1):105-12
9658195
Endocrinology. 2006 Oct;147(10):4831-42
16809442
J Biol Chem. 2001 Apr 27;276(17):14187-94
11297518
Science. 1995 Dec 1;270(5241):1491-4
7491495
Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W249-52
15215389
J Cell Biochem. 2003 Oct 1;90(2):315-26
14505348
J Steroid Biochem Mol Biol. 2000 Nov 30;74(5):311-7
11162939
Mol Cell Endocrinol. 2007 Aug 15;274(1-2):19-29
17597290
Breast Cancer Res. 2007;9(2):R25
17428314
J Biol Chem. 2000 Jun 16;275(24):18447-53
10749889
Nat Genet. 2006 Nov;38(11):1289-97
17013392
Nat Genet. 2000 May;25(1):25-9
10802651
J Biol Chem. 2001 Nov 9;276(45):42259-67
11533059
Cell. 1989 Feb 10;56(3):335-44
2644044
Proc Natl Acad Sci U S A. 2006 Aug 29;103(35):13162-7
16938840
J Biol Chem. 2000 Feb 25;275(8):5379-87
10681512
J Cell Biochem. 2007 Aug 1;101(5):1125-47
17520659
Oncogene. 2003 Aug 7;22(32):5011-20
12902984
Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1339-44
15677324
Oncogene. 2008 Feb 7;27(7):1019-32
17700529
Mol Cell Biol. 2003 Aug;23(16):5867-81
12897156