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Immortalization and transformation of human mammary epithelial cells by a tumor-derived Myc mutant

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Abstract

The Myc transcription factor is commonly dysregulated in many human cancers, including breast carcinomas. However, the precise role of Myc in the initiation and maintenance of malignancy is unclear. In this study we compared the ability of wild-type Myc (wt Myc) or Myc phosphorylation deficient mutants (T58A, S62A or T58A/S62A) to immortalize and transform human mammary epithelial cells (HMECs). All Myc constructs promoted cellular immortalization. As previously reported in other cells, the Myc T58A mutant tempered apoptotic responses and increased Myc protein stability in HMEC cells. More importantly, we now show that HMECs overexpressing the Myc T58A mutant acquire a unique cellular phenotype characterized by cell aggregation, detachment from the substrate and growth in liquid suspension. Coincident with these changes, the cells become anchorage-independent for growth in agarose. Previous studies have shown that wt Myc can collaborate with hTERT in inducing HMEC anchorage-independent growth. We have verified this observation and further shown that Myc T58A was a stronger facilitator of such co-transformation. Thus, our findings indicate that differences in Myc protein phosphorylation modulate its biological activity in human breast epithelial cells and specifically that the T58A mutation can facilitate both cellular immortalization and transformation. Finally, we used the isogenic cell lines generated in this study to identify a subset of genes whose expression is greatly altered during the transition from the immortal to the anchorage-independent states.

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Acknowledgements

This research was funded by Department of Defense Grant W81XWH-05-0259 (CT) and CA106400-03/5 from the NCI (RS).

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Author notes

  1. Gary L. Disbrow

    Present address: Department of Health and Human Services, Biomedical Advanced Research and Development Authority, Office of Preparedness and Response, Washington, DC, 20201, USA

Authors and Affiliations

  1. Department of Pathology, Georgetown University Medical School, 3900 Reservoir Road, NW, Washington, DC, 20057, USA

    Clare A. Thibodeaux, Xuefeng Liu, Gary L. Disbrow, Yiyu Zhang & Richard Schlegel

  2. Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical School, Washington, DC, 20057, USA

    Janice D. Rone & Bassem R. Haddad

  3. Department of Obstetrics and Gynecology, Lombardi Comprehensive Cancer Center, Georgetown University Medical School, Washington, DC, 20057, USA

    Bassem R. Haddad

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  1. Clare A. Thibodeaux
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Correspondence to Richard Schlegel.

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Thibodeaux, C.A., Liu, X., Disbrow, G.L. et al. Immortalization and transformation of human mammary epithelial cells by a tumor-derived Myc mutant. Breast Cancer Res Treat 116, 281–294 (2009). https://doi.org/10.1007/s10549-008-0127-x

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