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High-resolution mapping of molecular events associated with immortalization, transformation, and progression to breast cancer in the MCF10 model

  • Preclinical study
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Breast Cancer Research and Treatment Aims and scope Submit manuscript

Summary

Background

A comprehensive and consistent picture of the genetic changes that underlie breast cancer initiation, development, and progression remains unresolved. The MCF10 series of cell lines represents many steps in that progression. We performed high resolution mapping of the MCF10 series of cell lines to identify specific gene targets to elucidate the molecular correlates of immortalization, development, and progression of breast cancer at the level of individual genes.

Design

We evaluated the initial untransformed outgrowths (MCF-10MS and MCF-10A) with six transformed cell lines with benign proliferations (MCF-10AT1, MCF-10AT1kcl2), carcinoma in situ (MCF-10CA1h cl13), and invasive carcinoma (MCF-10CA1h cl2, MCF-10CA1a cl1, MCF-10CA1d cl1). Losses and gains of loci at 112 unique human genome sites were interrogated using the multiplex ligation-dependent probe amplification assay (MLPA).

Results

Cytogenetic alterations in the four benign progenitors that persisted in the CIS and invasive cell lines corresponded to gains and losses of genes by MLPA. MCF-10MS had only normal gene copies. The untransformed MCF-10A had cytogenetic gain of 5q13-qter with corresponding gains of the IL3, IL4 and IL12B genes at 5q31-q33; gain of distal 19q12-qter was reflected in gains in KLK3 and BAX gene loci at 19q13-q13.4. The observed genic gain of cMYC at 8q24.12 was not indicated by cytogenetics. The apparently balanced t(3;9) component of the t(3;9)(p13;p22)t(3;5)(p26;q31) resulted in complete loss of the CDKN2A and CDKN2B genes at 9p21. Additional clonal cytogenetic changes in the DCIS cell line (MCF-10A1h cl13) involving chromosomes 1, 3 and 10 persisted in the invasive progeny, with gain of corresponding genes at 1p13 (BCAR2, BCAR3, NRAS, TGFB2), at 3p12–13 (IL12A), and 3q21–27 (MME, PIK3CA, BCL6).

Conclusions

Our study adopted a comprehensive exploration of genetic changes using high resolution molecular probes applied to the MCF10 family of cell lines to identify individual genes in a continuum starting from normal breast epithelial cells and progressing through immortalization, transformation and invasive malignancy. Homozygous loss of CDKN2A and CDKN2B genes and gain of MYC were initiating immortalization events. Transformation and progression to malignancy event were marked by gains of IL13, VEGF, HRAS, TRAF2, and BCAS2, IL12A, and MME, respectively.

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Acknowledgements

Supported by in part by DOD Grants, DAMD DAMD17-00-1-0288 (MJW), DAMD17-02-1-0406 (MJW), and NIH Grants CA70923 (MJW), and DE15990 (MJW)

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Authors and Affiliations

  1. Department of Otolaryngology/Head and Neck Surgery, Henry Ford Hospital, Detroit, MI, USA

    Maria J. Worsham & Nivedita Tiwari

  2. Department of Clinical Genetics, VU Medical Center, Amsterdam, The Netherlands

    Gerard Pals & Rosalina van Spaendonk

  3. MRC Holland, Amsterdam, The Netherlands

    Jan P. Schouten

  4. Karmanos Cancer Center, Detroit, MI, USA

    Fred Miller

  5. Department of Pathology, George Washington University, Washington DC, Washington DC, USA

    Sandra R. Wolman

  6. Department of Otolaryngology/Head and Neck Research, Department of Pathology, Wayne State School of Medicine, Henry Ford Hospital 1 Ford Place, 1D, Detroit, MI, 48202, USA

    Maria J. Worsham

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  1. Maria J. Worsham
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  2. Gerard Pals
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Correspondence to Maria J. Worsham.

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Worsham, M.J., Pals, G., Schouten, J.P. et al. High-resolution mapping of molecular events associated with immortalization, transformation, and progression to breast cancer in the MCF10 model. Breast Cancer Res Treat 96, 177–186 (2006). https://doi.org/10.1007/s10549-005-9077-8

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