Human Melanoma cells over-express extracellular matrix 1 (ECM1) which is regulated by TFAP2C

Geeta Lal; Piedad Gómez-Contreras; Mikhail Kulak; George Woodfield; Thomas Bair; Frederick E Domann; Ronald J Weigel

doi:10.1371/journal.pone.0073953

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Human Melanoma cells over-express extracellular matrix 1 (ECM1) which is regulated by TFAP2C

Journal article

Open access

Peer reviewed

Human Melanoma cells over-express extracellular matrix 1 (ECM1) which is regulated by TFAP2C

Geeta Lal, Piedad Gómez-Contreras, Mikhail Kulak, George Woodfield, Thomas Bair, Frederick E Domann and Ronald J Weigel

PloS one, Vol.8(9), pp.e73953-e73953

2013

DOI: 10.1371/journal.pone.0073953

PMCID: PMC3759440

PMID: 24023917

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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.

Base Sequence

Extracellular Matrix Proteins - genetics

Humans

Transcription Initiation Site

Gene Expression Regulation, Neoplastic

Cell Line, Tumor

Molecular Sequence Data

Melanoma - pathology

Binding Sites

Cell Adhesion

Promoter Regions, Genetic - genetics

Transcription Factor AP-2 - metabolism

Details

Title: Subtitle: Human Melanoma cells over-express extracellular matrix 1 (ECM1) which is regulated by TFAP2C
Creators: Geeta Lal - Department of Surgery, University of Iowa, Iowa City, Iowa, United States of America
Piedad Gómez-Contreras
Mikhail Kulak
George Woodfield
Thomas Bair
Frederick E Domann
Ronald J Weigel
Resource Type: Journal article
Publication Details: PloS one, Vol.8(9), pp.e73953-e73953
DOI: 10.1371/journal.pone.0073953
PMID: 24023917
PMCID: PMC3759440
NLM abbreviation: PLoS One
ISSN: 1932-6203
eISSN: 1932-6203
Publisher: Public Library of Science; United States
Grant note: P30 ES005605 / NIEHS NIH HHS K08 CA151658 / NCI NIH HHS R01CA109294 / NCI NIH HHS NIH 1 K08 CA151658-01A1 / NCI NIH HHS R01 CA109294 / NCI NIH HHS
Language: English
Date published: 2013
Academic Unit: Molecular Physiology and Biophysics; Anatomy and Cell Biology; Stead Family Department of Pediatrics; Pathology; Surgery; Radiation Oncology; Biochemistry and Molecular Biology
Record Identifier: 9984047691002771

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