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Generation of MCF-7 cells with aggressive metastatic potential in vitro and in vivo

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

An Erratum to this article was published on 01 February 2016

Abstract

Epithelial-mesenchymal transition (EMT) is a cellular development program characterized by loss of cell adhesion and increased cell mobility. It is essential for numerous processes including metastasis. In this study we have generated “aggressive” MCF-7 breast cancer cells (MCF-7-EMT), which show significantly increased invasion in contrast to wild type MCF-7 (MCF-7 WT) cells. In addition, we have analyzed, whether these cell lines differ in their metastatic behavior in vivo and in expression of invasion and/or EMT-relevant genes. Invasive behavior of different human breast cancer cell lines was tested. “Aggressive” MCF-7 cells (MCF-7-EMT) were generated using coculture and mammosphere culture techniques. To analyze whether or not MCF-7-EMT cells in contrast to MCF-7 WT cells form metastases in vivo, we assessed metastases in a nude mouse model. mRNA expression profiles of MCF-7 WT cells and MCF-7-EMT cells were compared using the Affymetrix micro array technique. Expression of selected genes was validated using real-time PCR. In addition, protein expression of epithelial marker E-cadherin (CDH1) and mesenchymal markers N-cadherin (CDH2), Vimentin (VIM), and TWIST was compared. The breast cancer cell lines showed different invasive behavior from hardly any invasion to a stronger cell movement. Coculture with osteoblast-like MG63 cells led to significantly increased cell invasion rates. The highest increase was shown using MCF-7 WT cells. Generated MCF-7-EMT cells showed significantly increased invasion as compared to MCF-7 WT cells. In 8 of 10 mice bearing orthotopically growing MCF-7-EMT tumors, we could detect metastases in liver and lung. In mice bearing MCF-7 WT tumors (n = 10), no metastases were found. MCF-7 WT cells and MCF-7-EMT cells were different in expression of 325 genes. Forty-four of the most regulated 50 invasion and/or EMT-related genes were upregulated and 6 genes were downregulated in MCF-7-EMT cells. Protein expression of mesenchymal markers CDH2, VIM, and TWIST was clearly increased in MCF-7-EMT cells. Protein expression of epithelial marker CDH1 was clearly decreased. With the breast cancer cell lines, MCF-7-EMT and MCF-7 WT cells, we have an excellent model of cells for further studies of EMT and invasion in vitro and in vivo.

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Acknowledgments

We thank Sonja Blume, Renate Diedrich, and Matthias Läsche for their excellent technical assistance.

Conflict of interests

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Gynecology and Obstetrics, Georg-August-University, Robert-Koch-Street 40, 37075, Göttingen, Germany

    Elke Ziegler, Marie-Therese Hansen, Maike Haase, Günter Emons & Carsten Gründker

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  1. Elke Ziegler
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  2. Marie-Therese Hansen
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  3. Maike Haase
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  4. Günter Emons
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  5. Carsten Gründker
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Corresponding author

Correspondence to Carsten Gründker.

Additional information

Elke Ziegler and Marie-Therese Hansen have contributed equally to this work.

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Ziegler, E., Hansen, MT., Haase, M. et al. Generation of MCF-7 cells with aggressive metastatic potential in vitro and in vivo. Breast Cancer Res Treat 148, 269–277 (2014). https://doi.org/10.1007/s10549-014-3159-4

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  • DOI: https://doi.org/10.1007/s10549-014-3159-4

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