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    2D-DIGE analysis of phospho-enriched fractions from dasatinib-treated melanoma cell lines

    Eustace, Alex J. and Dowling, Paul and Henry, Michael and Doolan, Padraig and Meleady, Paula and Clynes, Martin and Crown, John and O'Donovan, Norma (2011) 2D-DIGE analysis of phospho-enriched fractions from dasatinib-treated melanoma cell lines. Journal of Proteomics, 74 (4). pp. 490-501. ISSN 1874-3919

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    Current therapeutic regimes for metastatic melanoma have failed to provide robust clinical responses. Dasatinib has shown anti-proliferative and anti-invasive effects in vitro; however, not all melanoma cells tested were sensitive to dasatinib. We used 2D-DIGE analysis of phospho-enriched fractions to identify phosphoproteins involved in regulating response to dasatinib in an isogenic pair of melanoma cell lines, one sensitive to dasatinib (WM-115) and the other resistant (WM-266-4). In WM-115 cells treated with dasatinib, 18 unique protein species with altered phosphorylation levels were detected. Dasatinib treatment of WM-266-4 cells resulted in phosphoprotein alterations to four unique protein species. Four phosphorylated forms of Annexin-A2 (ANXA2) were increased in WM-115 cells treated with dasatinib, whilst dasatinib treatment did not alter ANXA2 phosphoprotein levels in WM-266-4 cells. Immunoblotting confirmed that phosphorylation of ANXA2, on tyrosine residues, was increased in WM-115 cells treated with dasatinib. Subsequent knockdown of ANXA2 by siRNA significantly inhibited proliferation of WM-115 cells but did not significantly reduce proliferation of WM-266-4 cells. Therefore, ANXA2 plays a role in regulating proliferation in dasatinib-sensitive WM-115 cells and could potentially play a role in sensitivity to dasatinib in melanoma cells.

    Item Type: Article
    Additional Information: The definitive published version of this article is available at DOI: 10.1016/j.jprot.2010.12.011
    Keywords: 2D-DIGE; Phosphoproteomics; Dasatinib; Melanoma; Annexin-A2; ANXA2;
    Academic Unit: Faculty of Science and Engineering > Biology
    Item ID: 7335
    Identification Number:
    Depositing User: Paul Dowling
    Date Deposited: 12 Aug 2016 09:40
    Journal or Publication Title: Journal of Proteomics
    Publisher: Elsevier
    Refereed: Yes
    Use Licence: This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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