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Src activation of Stat3 is an independent requirement from NF-κB activation for constitutive IL-8 expression in human pancreatic adenocarcinoma cells

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Abstract

Human pancreatic tumors often overexpress the angiogenesis-promoting factor Interleukin 8 (IL-8), in part due to overexpression of NF-κB, a frequent occurrence in pancreatic adenocarcinoma. In this study, we demonstrate that reducing c-Src kinase activity, through either pharmacologic inhibition or small interfering RNA-targeted reduction of Src expression, significantly decreased IL-8 expression (P < 0.05) without affecting NF-κB-mediated transcription, but by decreasing phosphorylation of STAT3. To ascertain whether Src-mediated expression of IL-8 was dependent on STAT3, we used stable clones expressing a dominant-negative isoform of STAT3 that inhibits endogenous STAT3 phosphorylation and subsequent DNA binding and STAT3-mediated gene expression or a constitutively activated isoform of STAT3. IL-8 expression was significantly lower in clones expressing the dominant-negative isoform and significantly increased in clones expressing the activated isoform (P < 0.05 for both). Pharmacologic inhibition of NF-κB activity significantly reduced basal IL-8 expression and tumor necrosis factor-induced IL-8 expression (P < 0.05 for both), yet NF-κB activity was not dependent on Src. We therefore suggest that Src activation, through phosphorylation of␣STAT3, and NF-κB are all required for expression of IL-8 a critical angiogenic-promoting factor in pancreatic adenocarcinomas.

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Acknowledgements

We thank Elizabeth L. Hess, ELS (D), (The University of Texas M. D. Anderson Cancer Center) for editorial assistance and Dr. James Darnell (The Rockefeller University) for the STAT3-C and STAT3-Y705F plasmids. This work was supported by National Institute of Health (NIH) Grant T32 CA 09599 and the Eleanor B. Pillsbury Fellowship-University of Illinois Hospital (J.G.T.), NIH 2RO-1 CA65527 and NIH U54 CA 090810-01 and U.S. Department of Defense Grant PC020017 (G.E.G.), and the Lockton Foundation (M.J.G. and G.E.G.). M.J.G is the Lockton Fellow for Pancreatic Cancer Research.

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Authors and Affiliations

  1. Department of Cancer Biology, Unit 179, The University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX, 77030, USA

    Jose G. Trevino, Michael J. Gray, Steffan T. Nawrocki, Justin M. Summy, Donald P. Lesslie, David J. McConkey & Gary E. Gallick

  2. Department of Surgical Oncology, The University of Texas M. D. Anderson Cancer Center, Houston, TX, 77030, USA

    Jose G. Trevino, Donald P. Lesslie, Douglas B. Evans & Paul J. Chiao

  3. Department of Immunology, The University of Texas M. D. Anderson Cancer Center, Houston, TX, 77030, USA

    Stephanie S. Watowich

  4. Program in Cancer Biology, The University of Texas Graduate School of Biomedical Sciences, Houston, TX, USA

    Steffan T. Nawrocki, David J. McConkey & Gary E. Gallick

  5. Ariad Pharmaceuticals, Inc., Cambridge, MA, USA

    Tomi K. Sawyer & William C. Shakespeare

Authors
  1. Jose G. Trevino
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  2. Michael J. Gray
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  3. Steffan T. Nawrocki
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  4. Justin M. Summy
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  9. Stephanie S. Watowich
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  12. Gary E. Gallick
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Corresponding author

Correspondence to Gary E. Gallick.

Additional information

Jose G. Trevino and Michael J. Gray contributed equally to the work in this manuscript.

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Trevino, J.G., Gray, M.J., Nawrocki, S.T. et al. Src activation of Stat3 is an independent requirement from NF-κB activation for constitutive IL-8 expression in human pancreatic adenocarcinoma cells. Angiogenesis 9, 101–110 (2006). https://doi.org/10.1007/s10456-006-9038-9

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  • DOI: https://doi.org/10.1007/s10456-006-9038-9

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