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    Microcell-mediated chromosome transfer identifies EPB41L3 as a functional suppressor of epithelial ovarian cancers.

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    Authors
    Dafou, Dimitra
    Grun, Barbara
    Sinclair, Jonathan
    Lawrenson, Kate
    Benjamin, Elizabeth C.
    Hogdall, Estrid
    Kruger-Kjaer, Susanne
    Christensen, Lise
    Sowter, Heidi M. cc
    Al-Attar, Ahmed
    Edmondson, Richard
    Darby, Stephen
    Berchuck, Andrew
    Laird, Peter W.
    Pearce, C. Leigh
    Ramus, Susan J.
    Jacobs, Ian J.
    Gayther, Simon A.
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    Affiliation
    University College London, EGA Institute for Women's Health, Gynaecological Cancer Research Laboratories
    Issue Date
    2010-07
    
    Metadata
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    Abstract
    We used a functional complementation approach to identify tumor-suppressor genes and putative therapeutic targets for ovarian cancer. Microcell-mediated transfer of chromosome 18 in the ovarian cancer cell line TOV21G induced in vitro and in vivo neoplastic suppression. Gene expression microarray profiling in TOV21G(+18) hybrids identified 14 candidate genes on chromosome 18 that were significantly overexpressed and therefore associated with neoplastic suppression. Further analysis of messenger RNA and protein expression for these genes in additional ovarian cancer cell lines indicated that EPB41L3 (erythrocyte membrane protein band 4.1-like 3, alternative names DAL-1 and 4.1B) was a candidate ovarian cancer-suppressor gene. Immunoblot analysis showed that EPB41L3 was activated in TOV21G(+18) hybrids, expressed in normal ovarian epithelial cell lines, but was absent in 15 (78%) of 19 ovarian cancer cell lines. Using immunohistochemistry, 66% of 794 invasive ovarian tumors showed no EPB41L3 expression compared with only 24% of benign ovarian tumors and 0% of normal ovarian epithelial tissues. EPB41L3 was extensively methylated in ovarian cancer cell lines and primary ovarian tumors compared with normal tissues (P = .00004), suggesting this may be the mechanism of gene inactivation in ovarian cancers. Constitutive reexpression of EPB41L3 in a three-dimensional multicellular spheroid model of ovarian cancer caused significant growth suppression and induced apoptosis. Transmission and scanning electron microscopy demonstrated many similarities between EPB41L3-expressing cells and chromosome 18 donor-recipient hybrids, suggesting that EPB41L3 is the gene responsible for neoplastic suppression after chromosome 18 transfer. Finally, an inducible model of EPB41L3 expression in three-dimensional spheroids confirmed that reexpression of EPB41L3 induces extensive apoptotic cell death in ovarian cancers.
    Citation
    Microcell-mediated chromosome transfer identifies EPB41L3 as a functional suppressor of epithelial ovarian cancers. 2010, 12 (7):579-89 Neoplasia
    Journal
    Neoplasia (New York, N.Y.)
    URI
    http://hdl.handle.net/10545/252992
    PubMed ID
    20651987
    Type
    Article
    Language
    en
    ISSN
    1476-5586
    Collections
    Human Sciences Research Centre

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