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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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