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Adipose tissue derived stem cells differentiate into carcinoma-associated fibroblast-like cells under the influence of tumor derived factors

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Abstract

Background

Carcinoma-associated fibroblasts (CAF) are considered to contribute to tumor growth, invasion and metastasis. However, the cell type of origin remains unknown. Since human adipose tissue derived stem cells (hASCs) are locally adjacent to breast cancer cells and might directly interact with tumor cells, we investigated whether CAFs may originate from hASCs.

Methods

hASCs cultured under different conditions were quantified for the expression of alpha smooth muscle actin. ELISA was performed using the human TGFβ1, SDF-1α and CCL5 Quantikine Kit. The invasion potential of MDAMB231 cancer cells was evaluated using a Boyden chamber with filter inserts coated with Matrigel in 24-well dishes.

Results

We demonstrated that a significant percentage of hASCs differentiated into a CAF-like myofibroblastic phenotype (e.g. expression of alpha smooth muscle actin and tenascin-C) when exposed to conditioned medium from the human breast cancer lines MDAMB231 and MCF7. The conditioned medium from MDAMB231 and MCF7 contains significant amounts of transforming growth factor-beta 1 (TGFβ1) and the differentiation of hASCs towards CAFs is dependent on TGFβ1 signaling via Smad3 in hASCs. The induction of CAFs can be abolished using a neutralizing antibody to TGFβ1 as well as by pretreatment of the hASCs with SB431542, a TGFβ1 receptor kinase inhibitor. Additionally, we found that these hASC-derived CAF-like cells exhibit functional properties of CAFs, including the ability to promote tumor cell invasion in an in vitro invasion assay, as well as increased expression of stromal-cell derived factor 1 (SDF-1) and CCL5.

Conclusion

Our data suggest that hASCs are a source of CAFs which play an important role in the tumor invasion.

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Acknowledgements

This research was supported in part by the Department of Defense Breast Cancer Research Program W81XWH-08-1-0523 01 (to YHS) and by the Alliance of Cardiovascular Researchers (to EA).We thank Feras J Abdul Khalek and Christoph Beckmann for their technical assistance. The authors are also grateful to Dr. Sendurai Mani for helpful comments on this study.

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Authors and Affiliations

  1. Department of Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston, TX, 77030, USA

    Constantin Jotzu, Eckhard Alt, Gabriel Welte, Eswaran Devarajan, Srinivasalu Krishnappa, Severin Pinilla, Lilly Droll & Yao-Hua Song

  2. Department of Systems Biology and Gynecologic Medical Oncology, University of Texas M.D. Anderson Cancer Center, Houston, TX, 77030, USA

    Jie Li & Bryan T. Hennessy

  3. University of Texas, MD Anderson Cancer Center, 2SCR3.3024, 7435 Fannin Street, Box 0951, Houston, TX, 77054, USA

    Yao-Hua Song

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  1. Constantin Jotzu
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  2. Eckhard Alt
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  3. Gabriel Welte
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  8. Severin Pinilla
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  10. Yao-Hua Song
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Corresponding author

Correspondence to Yao-Hua Song.

Additional information

Constantin Jotzu and Eckhard Alt contributed equally to this work.

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Supplementary Figure 1

MCF7 conditioned medium stimulates the secretion of SDF-1α from hASCs. Protein levels of SDF-1α secreted from hASCs exposed to MCF7 conditioned medium over time (12–72 h) were measured by ELISA. The experiment was repeated three times. *P < 0.002 and **P < 0.0003 versus 12 h MCF7 CM. SDF-1α: stromal cell-derived factor 1 alpha; CM: conditioned medium. (PPT 105 kb)

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Jotzu, C., Alt, E., Welte, G. et al. Adipose tissue derived stem cells differentiate into carcinoma-associated fibroblast-like cells under the influence of tumor derived factors. Cellular Onc. 34, 55–67 (2011). https://doi.org/10.1007/s13402-011-0012-1

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