title: Human Melanoma cells over-express extracellular matrix 1 (ECM1) which is regulated by TFAP2C.
abstract: Extracellular matrix 1 (ECM1) is over-expressed in multiple epithelial malignancies. However, knowledge regarding the expression of ECM1 in melanomas and the mechanisms of ECM1 regulation is limited. In this study, we found that ECM1 is over-expressed in several melanoma cell lines, when compared to primary melanocytes, and furthermore, that ECM1 expression paralleled that of TFAP2C levels in multiple cell lines. Knockdown of TFAP2C in the A375 cell line with siRNA led to a reduction in ECM1 expression, and upregulation of TFAP2C with adenoviral vectors in the WM793 cell line resulted in ECM1 upregulation. Utilizing 5' RACE to identify transcription start sites (TSS) and luciferase reporter assays in the ECM1-overexpressing A375 cell line, we identified the minimal promoter region of human ECM1 and demonstrate that an approximately 100bp fragment upstream of the TSS containing a TATA box and binding sites for AP1, SP1 and Ets is sufficient for promoter activity. Chromatin immunoprecipitation and direct sequencing (ChIP-seq) for TFAP2C in the A375 cell line identified an AP2 regulatory region in the promoter of the ECM1 gene. Gelshift assays further confirmed binding of TFAP2C to this site. ECM1 knockdown reduces melanoma cell attachment and is consistent with findings that ECM1 overexpression has been associated with a poor prognosis. Our investigations show an as yet unrecognized role for TFAP2C in melanoma via its regulation of ECM1.
authors.affiliation: Department of Surgery, University of Iowa, Iowa City, Iowa, United States of America.
authors.forename: Geeta
authors.initials: G
authors.lastname: Lal
authors.forename: Piedad Gomez
authors.initials: PG
authors.lastname: Contreras
authors.forename: Mikhail
authors.initials: M
authors.lastname: Kulak
authors.forename: George
authors.initials: G
authors.lastname: Woodfield
authors.forename: Thomas
authors.initials: T
authors.lastname: Bair
authors.forename: Frederick E
authors.initials: FE
authors.lastname: Domann
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Weigel
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008545:Melanoma / Q000473:pathology*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D050656:Transcription Factor AP-2 / Q000378:metabolism*; D024363:Transcription Initiation Site
chemical_list: C490607:ECM1 protein, human; D016326:Extracellular Matrix Proteins; C493679:TFAP2C protein, human; D050656:Transcription Factor AP-2
references.citation: Hum Reprod. 2007 Jan;22(1):167-73
references.pmid: 16920726
references.citation: J Invest Dermatol. 2000 Apr;114(4):718-24
references.pmid: 10733679
references.citation: Int J Cancer. 2005 Jun 20;115(3):470-7
references.pmid: 15700319
references.citation: Ann Surg Oncol. 2009 Aug;16(8):2280-7
references.pmid: 19521735
references.citation: Gynecol Oncol. 2006 Mar;100(3):462-8
references.pmid: 16216317
references.citation: J Clin Oncol. 1998 Nov;16(11):3584-91
references.pmid: 9817279
references.citation: Biotechniques. 2009 Apr;46(4):265-74
references.pmid: 19450233
references.citation: Ann Surg. 2005 Sep;242(3):353-61; discussion 361-3
references.pmid: 16135921
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D885-90
references.pmid: 18940857
references.citation: Clin Cancer Res. 2009 Jun 1;15(11):3672-9
references.pmid: 19458056
references.citation: BMC Cancer. 2005 Jan 10;5:3
references.pmid: 15642117
references.citation: Cancer Invest. 2008 May;26(4):375-84
references.pmid: 18443958
references.citation: Gene. 1999 Jan 21;226(2):253-61
references.pmid: 9931498
references.citation: Cancer. 2006 Jun 15;106(12):2592-7
references.pmid: 16688775
references.citation: EMBO J. 2011 May 18;30(10):1990-2007
references.pmid: 21468029
references.citation: Mol Cancer Res. 2011 Jun;9(6):712-23
references.pmid: 21460181
references.citation: Nucleic Acids Res. 1999 Oct 15;27(20):4040-9
references.pmid: 10497269
references.citation: J Surg Res. 2007 Mar;138(1):71-8
references.pmid: 17187826
references.citation: Genomics. 1997 Nov 1;45(3):487-95
references.pmid: 9367673
references.citation: Cancer Lett. 2003 Oct 8;200(1):57-67
references.pmid: 14550953
references.citation: PLoS Genet. 2010 Sep 16;6(9):e1001122
references.pmid: 20862309
references.citation: Cancer Biol Ther. 2008 Jul;7(7):997-1003
references.pmid: 18698165
references.citation: FASEB J. 2001 Apr;15(6):988-94
references.pmid: 11292659
references.citation: Int J Cancer. 2007 May 15;120(10):2061-7
references.pmid: 17330235
references.citation: J Surg Res. 2008 Sep;149(1):62-8
references.pmid: 18374945
references.citation: Neoplasia. 2006 Jul;8(7):568-77
references.pmid: 16867219
references.citation: Nucleic Acids Res. 1994 Apr 25;22(8):1413-20
references.pmid: 8190633
references.citation: Matrix Biol. 1997 Nov;16(5):289-92
references.pmid: 9501329
references.citation: Neoplasma. 2012;59(4):409-15
references.pmid: 22489696
references.citation: Exp Mol Pathol. 1999 Apr;66(1):99-108
references.pmid: 10331969
references.citation: Med Oncol. 2011 Dec;28 Suppl 1:S318-25
references.pmid: 21128013