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EpCAM overexpression prolongs proliferative capacity of primary human breast epithelial cells and supports hyperplastic growth - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Molecular Cancer

, 12:56

First Online: 10 June 2013Received: 23 July 2012Accepted: 05 June 2013DOI: 10.1186-1476-4598-12-56

Cite this article as: Martowicz, A., Rainer, J., Lelong, J. et al. Mol Cancer 2013 12: 56. doi:10.1186-1476-4598-12-56

Abstract

IntroductionThe Epithelial Cell Adhesion Molecule EpCAM has been shown to be strongly expressed in human breast cancer and cancer stem cells and its overexpression has been supposed to support tumor progression and metastasis. However, effects of EpCAM overexpression on normal breast epithelial cells have never been studied before. Therefore, we analyzed effects of transient adenoviral overexpression of EpCAM on proliferation, migration and differentiation of primary human mammary epithelial cells HMECs.

MethodsHMECs were transfected by an adenoviral system for transient overexpression of EpCAM. Thereafter, changes in cell proliferation and migration were studied using a real time measurement system. Target gene expression was evaluated by transcriptome analysis in proliferating and polarized HMEC cultures. A Chicken Chorioallantoic Membrane CAM xenograft model was used to study effects on in vivo growth of HMECs.

ResultsEpCAM overexpression in HMECs did not significantly alter gene expression profile of proliferating or growth arrested cells. Proliferating HMECs displayed predominantly glycosylated EpCAM isoforms and were inhibited in cell proliferation and migration by upregulation of p27 and p53. HMECs with overexpression of EpCAM showed a down regulation of E-cadherin. Moreover, cells were more resistant to TGF-β1 induced growth arrest and maintained longer capacities to proliferate in vitro. EpCAM overexpressing HMECs xenografts in chicken embryos showed hyperplastic growth, lack of lumen formation and increased infiltrates of the chicken leukocytes.

ConclusionsEpCAM revealed oncogenic features in normal human breast cells by inducing resistance to TGF-β1-mediated growth arrest and supporting a cell phenotype with longer proliferative capacities in vitro. EpCAM overexpression resulted in hyperplastic growth in vivo. Thus, we suggest that EpCAM acts as a prosurvival factor counteracting terminal differentiation processes in normal mammary glands.

KeywordsEpCAM p53 Primary mammary epithelial cells CAM model Transforming growth factor beta AbbreviationsEpCAMEpithelial cell adhesion molecule

TGF-β1Transforming growth factor beta 1

CAMChorioallantoic membrane

HMECsHuman mammary epithelial cells

GFPGreen fluorescent protein

RTRoom temperature

RT-PCRReverse transcriptase-polymerase chain reaction

SA-β-galSenescence associated-beta galactosidase

MOIMultiplicity of infection

EMTEpithelial to mesenchymal transition.

Electronic supplementary materialThe online version of this article doi:10.1186-1476-4598-12-56 contains supplementary material, which is available to authorized users.

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Autor: Agnieszka Martowicz - Johannes Rainer - Julien Lelong - Gilbert Spizzo - Guenther Gastl - Gerold Untergasser

Fuente: https://link.springer.com/







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