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Multiple forms of BRMS1 are differentially expressed in the MCF10 isogenic breast cancer progression model

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Abstract

Clinical studies evaluating the mRNA expression level of the BRMS1 metastasis suppressor in the progression of breast cancer have not been consistent. The purpose of this study was to characterize endogenous BRMS1 mRNA and protein in a model of the progression of breast cancer. BRMS1 protein expression was evaluated in the genetically related MCF10 cell lines representing ‘normal’ breast epithelial cells (MCF10A), pre-malignant breast disease (MCF10AT), comedo ductal carcinoma in situ (MCF10DCIS.com), and metastatic carcinoma (MCF10CAa.1 and MCF10CAd.1α) with two antibodies that recognize distinct epitopes in the BRMS1 protein. Nuclear expression of the characteristic ~35 kDa BRMS1 protein was detected in all cell lines. Because BRMS1 was expressed in the metastatic MCF10 variants, the BRMS1 exons were sequenced to scan for possible genetic mutations. BRMS1 was wild-type with the exception of a synonymous T/C transition in exon 7. However, alternatively spliced variants were detected by RT-PCR. Two variants, BRMS1.v2 and BRMS1.v4 were only detected in the MCF10A and AT cell lines, while BRMS1 and BRMS1.v3 were detected in all lines. These results demonstrate that expression of the characteristic ~35 kDa BRMS1 protein is not sufficient to prevent metastasis. The differential expression of alternative splice variants suggests caution should be taken when evaluating BRMS1 mRNA in clinical samples.

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Abbreviations

BRMS1:

Breast cancer metastasis suppressor 1

ARID4A:

AT-Rich interactive domain 4A

ER:

Estrogen receptor

PR:

Progesterone receptor

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Acknowledgments

This work was supported by United States Public Health Service Grants CA87728 (to DRW) and F32CA113037 (to DRH) and a grant from the National Foundation for Cancer Research Center for Metastasis Research (to DRW). We thank Drs. Fred Miller and Herbert Soule (Karmanos Cancer Institute) for generously providing the MCF10 cell line series. We also thank Alka Mehta and Blake Moore for technical assistance and Dr. Monica Richert and Alexandra Silveira for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Pathology, University of Alabama at Birmingham, 1670 University Blvd., room VH-G019A, Birmingham, AL, 35294-0019, USA

    Douglas R. Hurst, Yi Xie, Mick D. Edmonds & Danny R. Welch

  2. Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL, USA

    Danny R. Welch

  3. Department of Pharmacology/Toxicology, University of Alabama at Birmingham, Birmingham, AL, USA

    Danny R. Welch

  4. Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, AL, USA

    Danny R. Welch

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  1. Douglas R. Hurst
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  2. Yi Xie
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  4. Danny R. Welch
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Correspondence to Douglas R. Hurst or Danny R. Welch.

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Hurst, D.R., Xie, Y., Edmonds, M.D. et al. Multiple forms of BRMS1 are differentially expressed in the MCF10 isogenic breast cancer progression model. Clin Exp Metastasis 26, 89–96 (2009). https://doi.org/10.1007/s10585-008-9216-9

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  • DOI: https://doi.org/10.1007/s10585-008-9216-9

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