The Influence of Breast Tumour-Derived Factors and Wnt Antagonism on the Transformation of Adipose-Derived Mesenchymal Stem Cells into Tumour-Associated Fibroblasts
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Within the tumour stroma, a heterogeneous population of cell types reciprocally regulates cell proliferation, which considerably affects the progression of the disease. In this study, using tumour conditioned medium (TCM) derived from breast tumour cell lines – MCF7 and MDA MB 231, we have demonstrated the differentiation of adipose-derived mesenchymal stem cells (ADSCs) into tumour-associated fibroblasts (TAFs). Since the Wnt signalling pathway is a key signalling pathway driving breast tumour growth, the effect of the Wnt antagonist secreted frizzled-related protein 4 (sFRP4) was also examined. The response of ADSCs to TCM and sFRP4 treatments was determined by using cell viability assay to determine the changes in ADSC viability, immunofluorescence for mesenchymal markers, glucose uptake assay, and glycolysis stress test using the Seahorse Extracellular Flux analyser to determine the glycolytic activity of ADSCs. ADSCs have been shown to acquire a hyper-proliferative state, significantly increasing their number upon short-term and long-term exposure to TCM. Changes have also been observed in the expression of key mesenchymal markers as well as in the metabolic state of ADSCs. SFRP4 significantly inhibited the differentiation of ADSCs into TAFs by reducing cell growth as well as mesenchymal marker expression (cell line-dependent). However, sFRP4 did not induce further significant changes to the altered metabolic phenotype of ADSCs following TCM exposure. Altogether, this study suggests that the breast tumour milieu may transform ADSCs into a tumour-supportive phenotype, which can be altered by Wnt antagonism, but is independent of metabolic changes.
KeywordsAdipose-derived mesenchymal stem cells Tumour-associated fibroblasts Tumour conditioned medium Differentiation Wnt signalling sFRP4
Adipose-derived mesenchymal stem cells
Secreted frizzled-related protein 4
Tumour conditioned medium
Transforming growth factor-beta
Extracellular acidification rates
Oxygen consumption rates
We acknowledge the research and technical support from the School of Biomedical Sciences and Curtin Health Innovation Research Institute, Curtin University, where the work was carried out.
MV conceptualised, performed all experiments, analysed data, and drafted the manuscript. KK assisted the experiments performed using Seahorse flux analyser, its data analysis, interpretation and critical revision of the manuscript. FA was involved with conceptualisation and critical revision of manuscript. RD, PN were involved with critical revision of the manuscript. AD was involved with conceptualisation, critical revision of the manuscript, and funding of the experiments. All authors have read and approved the final version of this manuscript.
MV is supported by scholarship from the Office of Research and Development, Faculty of Health sciences, Curtin University. MV would also like to acknowledge the contribution of an Australian Government Research Training Program Scholarship in supporting this research. AD is supported by strategic research funds from the School of Biomedical Sciences (Curtin University), Commercialisation Advisory Board of Curtin University, Cancer Council of Western Australia, and Actinogen Ltd., Perth, Western Australia.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflicts of interest.
Consent for Publication
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