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Palladin promotes invasion of pancreatic cancer cells by enhancing invadopodia formation in cancer-associated fibroblasts

Oncogene volume 33, pages 1265–1273 (2014)Cite this article

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Abstract

The stromal compartment surrounding epithelial-derived pancreatic tumors is thought to have a key role in the aggressive phenotype of this malignancy. Emerging evidence suggests that cancer-associated fibroblasts (CAFs), the most abundant cells in the stroma of pancreatic tumors, contribute to the tumor’s invasion, metastasis and resistance to therapy, but the precise molecular mechanisms that regulate CAFs behavior are poorly understood. In this study, we utilized immortalized human pancreatic CAFs to investigate molecular pathways that control the matrix-remodeling and invasion-promoting activity of CAFs. We showed previously that palladin, an actin-associated protein, is expressed at high levels in CAFs of pancreatic tumors and other solid tumors, and also in an immortalized line of human CAFs. In this study, we found that short-term exposure of CAFs to phorbol esters reduced the number of stress fibers and triggered the appearance of individual invadopodia and invadopodial rosettes in CAFs. Molecular analysis of invadopodia revealed that their composition resembled that of similar structures (that is, invadopodia and podosomes) described in other cell types. Pharmacological inhibition and small interfering RNA knockdown experiments demonstrated that protein kinase C, the small GTPase Cdc42 and palladin were necessary for the efficient assembly of invadopodia by CAFs. In addition, GTPase activity assays showed that palladin contributes to the activation of Cdc42. In mouse xenograft experiments using a mixture of CAFs and tumor cells, palladin expression in CAFs promoted the rapid growth and metastasis of human pancreatic tumor cells. Overall, these results indicate that high levels of palladin expression in CAFs enhance their ability to remodel the extracellular matrix by regulating the activity of Cdc42, which in turn promotes the assembly of matrix-degrading invadopodia in CAFs and tumor cell invasion. Together, these results identify a novel molecular signaling pathway that may provide new molecular targets for the inhibition of pancreatic cancer metastasis.

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Acknowledgements

We thank Dr Angela Mathison (Mayo Clinic College of Medicine) for assistance in generating the imPSCs (imPSC-C2 and imPSC-C3) and Gabriela Herrera for assistance with the AsPC-1 pancreatic tumor cells labeled with firefly luciferase. We also thank Dr Teresa Brentnall (University of Washington) for helpful discussions, and the Lineberger Comprehensive Cancer Center Animal Studies Core for assistance with xenograft experiments. This study was supported by grants from the NIH (GM081505 to CAO; CA161136 to SMG; DK52913 to RU), the NSF (MCB-1121365 to CAO), the Elsa U Pardee Foundation (CAO), the UNC University Cancer Research Fund (SMG), the CIHR (MOP-36332 to CAGM) and the Mayo Clinic Center for Cell Signaling in Gastroenterology (P30DK084567 to RU).

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Author notes

  1. S M Goicoechea, R García-Mata & A Valdivia

    Present address: 8Current address: Department of Biological Sciences, University of Toledo, Toledo, OH, USA.,

  2. H J Kim and C A Otey: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    S M Goicoechea, R García-Mata, J Staub, A Valdivia, L Sharek & C A Otey

  2. CIHR Group in Matrix Dynamics, Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada

    C G McCulloch

  3. Department of Surgical Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA

    R F Hwang

  4. Department of Biochemistry and Molecular Biology, Division of Gastroenterology and Hepatology, Epigenetics and Chromatin Dynamics Laboratory, Translational Epigenomics Program, Center for Individualized Medicine (CIM), Mayo Clinic, Rochester, MN, USA

    R Urrutia

  5. Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    J J Yeh

  6. Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    J J Yeh, H J Kim & C A Otey

  7. Department of Surgery, UNC School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    J J Yeh & H J Kim

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  1. S M Goicoechea
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Correspondence to S M Goicoechea.

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Goicoechea, S., García-Mata, R., Staub, J. et al. Palladin promotes invasion of pancreatic cancer cells by enhancing invadopodia formation in cancer-associated fibroblasts. Oncogene 33, 1265–1273 (2014). https://doi.org/10.1038/onc.2013.68

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