2010 SSAT PLENARY PRESENTATION Involvement of Osteopontin in the Matrix-Degrading and Proangiogenic Changes Mediated by Nicotine in Pancreatic Cancer Cells Melissa Lazar & Jennifer Sullivan & Galina Chipitsyna & Qiaoke Gong & Chee Y. Ng & Ahmed F. Salem & Tamer Aziz & Agnes Witkiewicz & David T. Denhardt & Charles J. Yeo & Hwyda A. Arafat Received: 30 April 2010 / Accepted: 18 August 2010 # 2010 The Society for Surgery of the Alimentary Tract Abstract Background Substantial evidence indicates that exposure to cigarette smoke is associated with an elevated risk of pancreatic ductal adenocarcinoma (PDA). However, the mechanisms underlying the effects of nicotine on the development or progression of PDA remain to be investigated. Previously, we showed that nicotine promotes the expression of osteopontin c (OPNc), an isoform of OPN protein that confers on cancer cells a migratory phenotype. In this study, we explored the potential prometastatic role of nicotine in PDA through studying its effect on the expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF) and evaluated the role of OPN in mediating these effects. Materials and Methods MMP-9 and VEGF mRNA and protein were analyzed in PDA cells treated with or without nicotine (3–300 nM). Transient transfection and luciferase-labeled promoter studies evaluated the effects of OPNc and OPN protein on the transcription and translation of MMP-9 and VEGF. Real-time PCR and immunohistochemistry were used to analyze the mRNA expression levels and localization of OPN, MMP-9, and VEGF proteins in matched invasive human PDA and surrounding nonmalignant tissues. Results and Discussion Nicotine significantly enhanced the expression of MMP-9 and VEGF mRNA and protein in PDA cells. Blocking OPN with siRNA or OPN antibody prevented the nicotine-mediated increase of both MMP-9 and VEGF. Transient transfection of OPNc gene in PDA cells or their treatment with recombinant OPN protein significantly (p <0.05) increased MMP-9 and VEGF mRNA expression levels and induced their promoter activities. In invasive PDA lesions, MMP-9 mRNA levels were significantly (p <0.005) higher in smokers vs. nonsmokers. VEGF protein co-localized with MMP-9 and OPN in the malignant ducts and correlated well with their higher levels in invasive PDA lesions. Conclusions Our data show for the first time that cigarette smoking and nicotine may contribute to PDA metastasis through inducing MMP-9 and VEGF and suggest that OPN plays a central role in mediating these effects. The presence of OPN as a downstream effector of nicotine that is capable of mediating its prometastatic effects in PDA cells is novel and could provide a unique therapeutic target to control pancreatic cancer aggressiveness, especially in the cigarette-smoking population. Keywords Pancreatic cancer . Nicotine . Osteopontin . VEGF . MMP-9 . Angiogenesis . Metastasis Introduction Pancreatic ductal adenocarcinoma (PDA) is an extremely aggressive cancer, and currently, there are no methods for early detection. At the time of diagnosis, more than 85% of the tumors have infiltrated into adjacent organs or have metasta- sized, and the overall 5-year survival rate is <5%. 1 Given the grim prognosis of this disease, it is essential to understand the mechanisms that underlie PDA aggressiveness in order to design more effective therapies. Exposure to cigarette smoke has been shown to be strongly associated with an elevated risk of PDA. Smokers have at least twofold increase in the risk of developing PDA, 2 and it is Presented as a plenary presentation at the 51st Annual Meeting of the Society for Surgery of the Alimentary Tract, May 1–5, 2010 and an oral presentation at the SSAT Residents and Fellows Research Conference, May 1, 2010, New Orleans, LA. M. Lazar : J. Sullivan : G. Chipitsyna : Q. Gong : C. Y. Ng : A. F. Salem : T. Aziz : A. Witkiewicz : D. T. Denhardt : C. J. Yeo : H. A. Arafat (*) Department of Surgery, Jefferson Pancreatic, Biliary and Related Cancer Center, Thomas Jefferson University, 1015 Walnut Street, Philadelphia, PA 19107, USA e-mail: hwyda.arafat@jefferson.edu J Gastrointest Surg DOI 10.1007/s11605-010-1338-0