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

Dimitra Dafou; Barbara Grun; John Sinclair; Kate Lawrenson; Elizabeth C Benjamin; Estrid Vilma Solyom Høgdall; Susanne Kruger-Kjaer; Lise Christensen; Heidi M Sowter; Ahmed Al-Attar; Richard Edmondson; Stephen Darby; Andrew Berchuck; Peter W Laird; C Leigh Pearce; Susan J Ramus; Ian J Jacobs; Simon A Gayther

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

Dimitra Dafou, Barbara Grun, John Sinclair, Kate Lawrenson, Elizabeth C Benjamin, Estrid Vilma Solyom Høgdall, Susanne Kruger-Kjaer, Lise Christensen, Heidi M Sowter, Ahmed Al-Attar, Richard Edmondson, Stephen Darby, Andrew Berchuck, Peter W Laird, C Leigh Pearce, Susan J Ramus, Ian J Jacobs, Simon A Gayther

35 Citations (Scopus)

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 EPB47 L3 (erythrocyte membrane protein band 4.1-like 3, alternative names DAL-1 and 4.1 B) was a candidate ovarian cancer-suppressor gene. Immunoblot analysis showed that EPB41'L3 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.

Original language	English
Journal	NeoPlasia
Volume	12
Issue number	7
Pages (from-to)	579-89
Number of pages	11
ISSN	1522-8002
Publication status	Published - 1 Jul 2010

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Dafou, D., Grun, B., Sinclair, J., Lawrenson, K., Benjamin, E. C., Høgdall, E. V. S., Kruger-Kjaer, S., Christensen, L., Sowter, H. M., Al-Attar, A., Edmondson, R., Darby, S., Berchuck, A., Laird, P. W., Pearce, C. L., Ramus, S. J., Jacobs, I. J., & Gayther, S. A. (2010). Microcell-mediated chromosome transfer identifies EPB41L3 as a functional suppressor of epithelial ovarian cancers. NeoPlasia, 12(7), 579-89.

Dafou, D, Grun, B, Sinclair, J, Lawrenson, K, Benjamin, EC, Høgdall, EVS, Kruger-Kjaer, S, Christensen, L, Sowter, HM, Al-Attar, A, Edmondson, R, Darby, S, Berchuck, A, Laird, PW, Pearce, CL, Ramus, SJ, Jacobs, IJ & Gayther, SA 2010, 'Microcell-mediated chromosome transfer identifies EPB41L3 as a functional suppressor of epithelial ovarian cancers', NeoPlasia, vol. 12, no. 7, pp. 579-89.

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title = "Microcell-mediated chromosome transfer identifies EPB41L3 as a functional suppressor of epithelial ovarian cancers",

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 EPB47 L3 (erythrocyte membrane protein band 4.1-like 3, alternative names DAL-1 and 4.1 B) was a candidate ovarian cancer-suppressor gene. Immunoblot analysis showed that EPB41'L3 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.",

author = "Dimitra Dafou and Barbara Grun and John Sinclair and Kate Lawrenson and Benjamin, {Elizabeth C} and H{\o}gdall, {Estrid Vilma Solyom} and Susanne Kruger-Kjaer and Lise Christensen and Sowter, {Heidi M} and Ahmed Al-Attar and Richard Edmondson and Stephen Darby and Andrew Berchuck and Laird, {Peter W} and Pearce, {C Leigh} and Ramus, {Susan J} and Jacobs, {Ian J} and Gayther, {Simon A}",

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pages = "579--89",

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T1 - Microcell-mediated chromosome transfer identifies EPB41L3 as a functional suppressor of epithelial ovarian cancers

AU - Dafou, Dimitra

AU - Grun, Barbara

AU - Sinclair, John

AU - Lawrenson, Kate

AU - Benjamin, Elizabeth C

AU - Høgdall, Estrid Vilma Solyom

AU - Kruger-Kjaer, Susanne

AU - Christensen, Lise

AU - Sowter, Heidi M

AU - Al-Attar, Ahmed

AU - Edmondson, Richard

AU - Darby, Stephen

AU - Berchuck, Andrew

AU - Laird, Peter W

AU - Pearce, C Leigh

AU - Ramus, Susan J

AU - Jacobs, Ian J

AU - Gayther, Simon A

PY - 2010/7/1

Y1 - 2010/7/1

N2 - 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 EPB47 L3 (erythrocyte membrane protein band 4.1-like 3, alternative names DAL-1 and 4.1 B) was a candidate ovarian cancer-suppressor gene. Immunoblot analysis showed that EPB41'L3 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.

AB - 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 EPB47 L3 (erythrocyte membrane protein band 4.1-like 3, alternative names DAL-1 and 4.1 B) was a candidate ovarian cancer-suppressor gene. Immunoblot analysis showed that EPB41'L3 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.

M3 - Journal article

SN - 1522-8002

VL - 12

SP - 579

EP - 589

JO - Neoplasia (United States)

JF - Neoplasia (United States)

IS - 7

ER -

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

Abstract

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