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Synergistic effect between EGF and TGF-β1 in inducing oncogenic properties of intestinal epithelial cells

Oncogene volume 27, pages 2626–2634 (2008)Cite this article

Abstract

Transforming growth factor (TGF)-β1 has a biphasic effect on rat intestinal epithelial (RIE) cells. By itself, TGF-β1 functions as a tumor suppressor by inhibiting the growth, migration and invasion of RIE cells. We show in this study that in conjunction with epidermal growth factor (EGF), TGF-β1 helped to augment migration, invasion and anchorage-independent growth (AIG) compared to that by EGF alone. EGF plus TGF-β1 induced a dramatic morphological change characteristic of epithelial–mesenchymal transition (EMT). The mechanism for this enhanced effect of TGF-β1 and EGF on oncogenic properties was explored by analysis of EGF- and TGF-β1-mediated signaling pathways and complementary DNA arrays. TGF-β1 augmented EGF-mediated signaling of mitogen-activated protein kinase (MAPK) and AKT by enhancing and prolonging the activation of the former and prolonging the activation of the latter. Inhibition of MAPK, but not phosphoinositide-3 kinase (PI3K), abolished TGF-β1 plus EGF-induced EMT and downregulation of E-cadherin at mRNA and protein levels. By contrast, cell migration and invasion were sensitive to inhibition of either MAPK or PI3 kinase. TGF-β1 plus EGF-induced AIG was significantly more resistant to inhibition of PI3K and MAPK compared to that induced by EGF alone. EGF and TGF-β1 synergistically induced the expression of a series of proteases including matrix metalloproteinase (MMP) 1 (collagenase), MMP3, MMP9, MMP10, MMP14 and cathepsin. Among them, the expression of MMP1, MMP3, MMP9 and MMP10 was MAPK dependent. Inhibition of the MMPs or cathepsin significantly blocked EGF plus TGF-β1-induced invasion, but had no effect on colony formation. Phospholipase C (PLC) and Cox2 induced by EGF plus TGF-β1 also played a significant role in invasion, whereas PLC was also important for colony formation. Our study reveals specific signaling functions and induction of genes differentially required for enhanced effect of EGF- and TGF-β1-induced oncogenic properties, and helps to explain the tumor-promoting effect of TGF-β1 in human cancer with elevated expression or activation of TGF-β1 and receptor protein tyrosine kinases.

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Acknowledgements

We thank Dr J Cheng of University of South Florida for the gift of AKT inhibitor AP1-2 and Dr Weizhou Zhang for artwork of the figures. We apologize to our many colleagues for not being able to cite their studies due to space limitation. This work was supported by NIH Grants CA29339 and CA55054 and DOD Grant DAMD 17-02-01-0504/BC11101.

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Author notes

  1. S Uttamsingh

    Present address: 5Current address: AMGEN, Clinical Research Management, Oncology Therapeutics, South San Francisco, CA, USA,

  2. K T Nguyen

    Present address: 6Current address: Division of General Surgery, Department of Surgery, University of Michigan, Ann Arbor, MI, USA,

  3. M Bhanot

    Present address: 7Current address: The Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY, USA,

  4. S Uttamsingh, X Bao and K T Nguyen: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Microbiology, Mount Sinai School of Medicine, New York, NY, USA

    S Uttamsingh, X Bao, K T Nguyen, M Bhanot, J Gong, J L-K Chan & L-H Wang

  2. Center for Advanced Biotechnology and Medicine and Susan Lehman Cullman Laboratory for Cancer Research, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, NJ, USA

    F Liu

  3. Department of Pharmacology and System Therapeutics, Mount Sinai School of Medicine, New York, NY, USA

    T T Chu

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Correspondence to L-H Wang.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Uttamsingh, S., Bao, X., Nguyen, K. et al. Synergistic effect between EGF and TGF-β1 in inducing oncogenic properties of intestinal epithelial cells. Oncogene 27, 2626–2634 (2008). https://doi.org/10.1038/sj.onc.1210915

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