Protein kinase D1 promotes anchorage-independent growth, invasion, and angiogenesis by human pancreatic cancer cells

Authors

  • Nobuo Ochi,

    1. Department of Gastroenterology, Hepatology and Nutrition, MD Anderson Cancer Center, The University of Texas, Houston, Texas
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  • Suebpong Tanasanvimon,

    1. Department of Gastroenterology, Hepatology and Nutrition, MD Anderson Cancer Center, The University of Texas, Houston, Texas
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  • Yoichi Matsuo,

    1. Department of Gastroenterology, Hepatology and Nutrition, MD Anderson Cancer Center, The University of Texas, Houston, Texas
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  • Zhimin Tong,

    1. Department of Gastroenterology, Hepatology and Nutrition, MD Anderson Cancer Center, The University of Texas, Houston, Texas
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  • Bokyung Sung,

    1. Department of Experimental Therapeutics, MD Anderson Cancer Center, The University of Texas, Houston, Texas
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  • Bharat B. Aggarwal,

    1. Department of Experimental Therapeutics, MD Anderson Cancer Center, The University of Texas, Houston, Texas
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  • James Sinnett-Smith,

    1. Division of Digestive Diseases and CURE, Digestive Diseases Research Center, David Geffen School of Medicine at UCLA, Los Angeles, California
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  • Enrique Rozengurt,

    Corresponding author
    1. Division of Digestive Diseases and CURE, Digestive Diseases Research Center, David Geffen School of Medicine at UCLA, Los Angeles, California
    • Division of Digestive Diseases, Department of Medicine, 900 Veteran Avenue, Warren Hall Room 11-124, David Geffen School of Medicine at UCLA, Los Angeles, CA 90095.
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  • Sushovan Guha

    Corresponding author
    1. Department of Gastroenterology, Hepatology and Nutrition, MD Anderson Cancer Center, The University of Texas, Houston, Texas
    • Department of Gastroenterology, Hepatology, and Nutrition, The UT MD Anderson Cancer Center, Unit 1466, 1515 Holcombe Boulevard, Houston, TX 77030.
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Abstract

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal diseases. Novel molecularly targeted therapies are urgently needed. Here, we extended our studies on the role of protein kinase D1 (PKD1) in PDAC cell lines. Given that Panc-1 express moderate levels of PKD1, we used retroviral-mediated gene transfer to create a Panc-1 derivative that stably over-expresses PKD1 (Panc-1-PKD1). Reciprocally, we used shRNA targeting PKD1 in Panc-28 to produce a PKD1 under-expressing Panc-28 derivative (Panc-28-shPKD1). Our results demonstrate that Panc-1-PKD1 cells exhibit significantly increased anchorage-independent growth in soft agar and increased in vitro invasion compared with Panc-1-mock. Reciprocally, Panc-28-shPKD1 cells show a significant decrease in anchorage-independent growth and invasiveness, as compared with Panc-28-mock cells. The selective PKD family inhibitor CRT0066101 markedly decreased colony-forming ability and invasiveness by either Panc-1-PKD1 or Panc-28-mock cells. Secretion of the pro-angiogenic factors vascular endothelial growth factor (VEGF) and CXC chemokines (CXCL8) was significantly elevated by PKD1 over-expression in Panc-1 cells and reduced either by depletion of PKD1 via shRNA in Panc-28 cells or by addition of CRT0066101 to either Panc-1-PKD1 or Panc-28-mock cells. Furthermore, human umbilical vein endothelial cell (HUVEC) tube formation was significantly enhanced by co-culture with Panc-1-PKD1 compared with Panc-1-mock in an angiogenesis assay in vitro. Conversely, PKD1 depletion in Panc-28 cells decreased their ability to induce endotube formation by HUVECs. PDAC-induced angiogenesis in vitro and in vivo was markedly inhibited by CRT0066101. Our results lend further support to the hypothesis that PKD family members provide a novel target for PDAC therapy. J. Cell. Physiol. 226: 1074–1085, 2011. © 2010 Wiley-Liss, Inc.

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