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A Novel 3-Dimensional Co-culture Method Reveals a Partial Mesenchymal to Epithelial Transition in Breast Cancer Cells Induced by Adipocytes

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Journal of Mammary Gland Biology and Neoplasia Aims and scope Submit manuscript

Abstract

Cancer metastases are accountable for almost 90% of all human cancer related deaths including from breast cancer (BC). Adipocytes can alter the tumor microenvironment, which can promote metastasis by inducing an epithelial-to-mesenchymal transition (EMT) in BC cells. However, the role of adipocytes during the mesenchymal-to-epithelial transition (MET), that can be important in metastasis, is not clear. To understand the effect of adipocytes on the BC progression, there is a requirement for a better in vitro 3-dimensional (3D) co-culture system that mimics the breast tissue and allows for more accurate analysis of EMT and MET. We developed a co-culture system to analyze the relationship of BC cells grown in a 3D culture with adipocytes. We found that adipocytes and adipocyte-derived conditioned media, but not pre-adipocytes, caused the mesenchymal MDA-MB-231 and Hs578t cells to form significantly more epithelial-like structures when compared to the typical stellate colonies formed in control 3D cultures. SUM159 cells and MCF7 cells had a less dramatic shift as they normally have more epithelial-like structure in 3D culture. Biomarker expression analysis revealed that adipocytes only induced a partial MET with proliferation unaffected. In addition, adipocytes had reduced lipid droplet size when co-cultured with BC cells. Thus, we found that physical interaction with adipocytes and ECM changes the mesenchymal phenotype of BC cells in a manner that could promote secondary tumor formation.

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Fig. 1: Schematic representation of the 3D co-culture system.
Fig. 2: Adipocytes alter the characteristic morphology of mesenchymal but not epithelial BC cell lines grown in 3D.
Fig. 3: Adipocytes partially enhance MET of mesenchymal BC cells in 3D culture with little to no effect on epithelial BC cells.
Fig. 4: Adipocytes increase CD24 in MDA-MB-21 cells with no effect on CD44 or the Ki67 proliferation marker in any cell line.
Fig. 5: Pre-adipocytes have a partial effect on morphology of BC cells.
Fig. 6: Pre-adipocytes have no effect on MET or on stemness markers of BC cells.
Fig. 7: Co-culture with BC cells decreases the size but not number of LD present in mature adipocytes. a

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Funding

Funding provided to SLC by the Memorial University of Newfoundland (210525), a New Investigator Award from the Beatrice Hunter Cancer Research Institute (207939), and the Cancer Research Society (22130). An NSERC Discovery Grant to AVP provided funding for optimization of the 3D culture model (RGPIN-2017-3977). NKP was supported by a trainee award and a Skills Acquisition Program award from the Beatrice Hunter Cancer Research Institute with funds provided by The Terry Fox Strategic Health Research Training Program in Cancer Research at CIHR and by Memorial University of Newfoundland.

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Authors and Affiliations

  1. Memorial University of Newfoundland, 232 Elizabeth Ave, St. Johns, NL, A1B 3X9, Canada

    Nikitha K. Pallegar, Chantae J. Garland & Sherri L. Christian

  2. Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada

    Mathepan Mahendralingam & Alicia M. Viloria-Petit

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Correspondence to Alicia M. Viloria-Petit or Sherri L. Christian.

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Pallegar, N.K., Garland, C.J., Mahendralingam, M. et al. A Novel 3-Dimensional Co-culture Method Reveals a Partial Mesenchymal to Epithelial Transition in Breast Cancer Cells Induced by Adipocytes. J Mammary Gland Biol Neoplasia 24, 85–97 (2019). https://doi.org/10.1007/s10911-018-9420-4

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