title: Transcriptomic and proteomic profiling of KEAP1 disrupted and sulforaphane-treated human breast epithelial cells reveals common expression profiles.
abstract: Sulforaphane (SFN), an isothiocyanate found in cruciferous vegetables, is a potent inhibitor of experimental mammary carcinogenesis and may be an effective, safe chemopreventive agent for use in humans. SFN acts in part on the Keap1/Nrf2 pathway to regulate a battery of cytoprotective genes. In this study, transcriptomic and proteomic changes in the estrogen receptor negative, non-tumorigenic human breast epithelial MCF10A cell line were analyzed following SFN treatment or KEAP1 knockdown with siRNA using microarray and stable isotopic labeling with amino acids in culture (SILAC), respectively. Changes in selected transcripts and proteins were confirmed by PCR and Western blot in MCF10A and MCF12A cells. There was strong correlation between the transcriptomic and proteomic responses in both the SFN treatment (R = 0.679, P < 0.05) and KEAP1 knockdown (R = 0.853, P < 0.05) experiments. Common pathways for SFN treatment and KEAP1 knockdown were xenobiotic metabolism and antioxidants, glutathione metabolism, carbohydrate metabolism, and NADH/NADPH regeneration. Moreover, these pathways were most prominent in both the transcriptomic and the proteomic analyses. The aldo-keto reductase family members, AKR1B10, AKR1C1, AKR1C2 and AKR1C3, as well as NQO1 and ALDH3A1, were highly upregulated at both the transcriptomic and the proteomic levels. Collectively, these studies served to identify potential biomarkers that can be used in clinical trials to investigate the initial pharmacodynamic action of SFN in the breast.
authors: Department of Biochemistry and Molecular Biology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA.
authors: Abena S
authors: AS
authors: Agyeman
authors: Raghothama
authors: R
authors: Chaerkady
authors: Patrick G
authors: PG
authors: Shaw
authors: Nancy E
authors: NE
authors: Davidson
authors: Kala
authors: K
authors: Visvanathan
authors: Akhilesh
authors: A
authors: Pandey
authors: Thomas W
authors: TW
authors: Kensler
references: J Nutr. 2005 Aug;135(8):1865-72
references: 16046710
references: Cancer Res. 2004 Oct 15;64(20):7610-7
references: 15492289
references: Hum Mol Genet. 2007 May 15;16(10):1188-200
references: 17409198
references: Biochem Biophys Res Commun. 1997 Jul 18;236(2):313-22
references: 9240432
references: Cancer Res. 2002 Sep 15;62(18):5196-203
references: 12234984
references: Cancer Prev Res (Phila). 2011 Mar;4(3):384-95
references: 21372038
references: Cancer Res. 2000 Feb 15;60(4):936-43
references: 10706108
references: Cancer Epidemiol Biomarkers Prev. 2005 Nov;14(11 Pt 1):2605-13
references: 16284385
references: Toxicol Sci. 2010 Aug;116(2):422-32
references: 20442190
references: J Biol Chem. 2008 Feb 8;283(6):3418-3423
references: 18056116
references: Clin Chim Acta. 2002 Feb;316(1-2):43-53
references: 11750273
references: Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3147-50
references: 8159717
references: Toxicol Lett. 2004 Nov 28;153(3):343-55
references: 15454310
references: Nutr Cancer. 2006;55(1):53-62
references: 16965241
references: J Proteomics. 2010 Jun 16;73(8):1612-31
references: 20399915
references: Nat Protoc. 2008;3(3):505-16
references: 18323819
references: Carcinogenesis. 2009 Jun;30(6):1024-31
references: 19386581
references: Carcinogenesis. 2009 Sep;30(9):1571-80
references: 19608619
references: BMC Cancer. 2003 Mar 22;3:9
references: 12659654
references: Carcinogenesis. 2007 Jul;28(7):1485-90
references: 17347138
references: Acta Pharmacol Sin. 2007 Sep;28(9):1343-54
references: 17723168
references: Ann Intern Med. 2009 Nov 17;151(10):703-15, W-226-35
references: 19920271
references: Proc Natl Acad Sci U S A. 1997 Sep 16;94(19):10367-72
references: 9294217
references: Am J Respir Cell Mol Biol. 2006 Dec;35(6):639-50
references: 16794261
references: J Nutr. 2007 Jul;137(7):1718-24
references: 17585021
references: Biochem J. 2006 Oct 1;399(1):101-9
references: 16787387
references: Cancer Res. 2007 Oct 15;67(20):9937-44
references: 17942926
references: Cell Stem Cell. 2007 Nov;1(5):555-67
references: 18371393
references: J Pharmacol Exp Ther. 2004 Jul;310(1):263-71
references: 14988420
references: Chem Biol Interact. 2011 May 30;191(1-3):185-91
references: 21277289
references: Biochem J. 2000 Oct 1;351(Pt 1):67-77
references: 10998348
references: Cancer Lett. 2006 Nov 18;243(2):170-92
references: 16516379
references: Cancer Res. 1990 Sep 15;50(18):6075-86
references: 1975513
references: Clin Cancer Res. 2005 Mar 1;11(5):1687-90
references: 15755988
references: Proc Natl Acad Sci U S A. 2005 May 17;102(20):7280-7285A
references: 15883370
references: CA Cancer J Clin. 2010 Sep-Oct;60(5):277-300
references: 20610543
references: Stem Cells. 2009 Jul;27(7):1487-95
references: 19544456
references: Clin Cancer Res. 2010 May 1;16(9):2580-90
references: 20388854
references: Proteome Sci. 2010 Dec 16;8:67
references: 21159204
references: Proc Natl Acad Sci U S A. 2008 May 13;105(19):6846-51
references: 18474869
references: Biopharm Drug Dispos. 2006 Dec;27(9):407-20
references: 16952200
references: J Biol Chem. 2010 Jul 16;285(29):22576-91
references: 20452972
references: Methods. 2001 Dec;25(4):402-8
references: 11846609
references: Int J Cancer. 1995 Sep 15;62(6):749-54
references: 7558425
references: Cancer Lett. 2006 Feb 28;233(2):208-18
references: 16520150
references: Med Sci Monit. 2006 Dec;12(12):CR525-529
references: 17136009
references: Cancer Res. 2008 Jul 1;68(13):5487-91
references: 18593952
references: Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2394-8
references: 1549602
references: Cell Mol Life Sci. 2007 May;64(9):1105-27
references: 17396224
references: J Biol Chem. 1992 Jul 25;267(21):15097-104
references: 1340765
references: Biochem Biophys Res Commun. 2011 Feb 11;405(2):173-9
references: 21216231
references: J Biol Chem. 2003 Mar 7;278(10):8135-45
references: 12506115
references: Prostate. 2009 Feb 1;69(2):181-90
references: 18973173
references: Anticancer Drugs. 2002 Sep;13(8):859-68
references: 12394272
references: Cancer Res. 2001 Aug 15;61(16):6120-30
references: 11507062
references: Int J Cancer. 1992 Feb 1;50(3):463-73
references: 1370949
references: Trends Biochem Sci. 2009 Apr;34(4):176-88
references: 19321346
references: Biochem Biophys Res Commun. 1996 Nov 12;228(2):474-9
references: 8920937
references: Breast Cancer (Auckl). 2010 May 20;4:35-41
references: 20697531