Antiprotease strategy in combination with gemcitabine as a putative novel therapeutic approach for pancreatic cancer treatment

none11Introduction: gabexate mesylate (gm), a protease inhibitor, has been shown to exert a significant antitumoral activity in pancreatic cancer (PC) cells. Aim: To evaluate whether pre-treatment with gm could improve PC cells response toward gemcitabine (gem). Methods: Studies have been performed on PanC-1, SW1990 PC cells and ea.hy926 endothelial cells. Treatment consisted of gm 50 µg/ml for 24 hrs, followed by 24 hrs of treatment with gem 6 µg/ml; 24 hrs of gm 50 µg/ml followed by 24 hrs in culture medium; 24 hrs of gem 6 µg/ml followed by 24 hrs in culture medium. aspects studied included: cell viability (mTT assay), cell invasiveness and migration (Boyden chambers chemoinvasion and chemotaxis assay), angiogenesis (endothelial tube formation assay), nf-kB activation (nf-kB luciferase assay, Western Blotting,), vegf and il-8 levels (eliSa), mmP-2 and mmP-9 activity (gelatin zymography), Ras signalling (Western Blotting). results: gm significantly enhanced gem anti-invasive and anti-angiogenic efficacy by inhibition of gem-induced nf-kB activation (P< 0.001). This phenomenon occurred by the block of ikBalpha degradation that, in turn, prevented Rela/p65 nuclear translocation (P<0.001) and resulted in down-regulation of the nf-kB products mmP-2 and mmP-9, vegf and il-8 (P< 0.001). drugs in combination also inhibited eRK1/2 and aKT activation, two pathways targeting nf-kB, by increased expression of their endogenous inhibitors RKiP and PTen, respectively. gm did not enhance the gem action on cell viability after 24 hrs. Conclusion: These in vitro data indicate that gm pretreatment significantly improves gem anti-invasive and anti-angiogenic efficacy in PC cells, in part by nf-kB inhibition.Pancreatology is the official journal of the International Association of Pancreatology (IAP), the European Pancreatic Club (EPC) and several national societies and study groups around the world. Dedicated to the understanding and treatment of exocrine as well as endocrine pancreatic disease, this multidisciplinary periodical publishes original basic, translational and clinical pancreatic research from a range of fields including gastroenterology, oncology, surgery, pharmacology, cellular and molecular biology as well as endocrinology, immunology and epidemiology. Readers can expect to gain new insights into pancreatic physiology and into the pathogenesis, diagnosis, therapeutic approaches and prognosis of pancreatic diseases. The journal features original articles, case reports, consensus guidelines and topical, cutting edge reviews, thus representing a source of valuable, novel information for clinical and basic researchers alike.mixedM Macchini; G Brandi; S Tavolari; T Guarnieri; M Di Marco; P Paterini; F De Rosa; S Di Girolamo; S Vecchiarelli; A Papi; G BiascoM Macchini; G Brandi; S Tavolari; T Guarnieri; M Di Marco; P Paterini; F De Rosa; S Di Girolamo; S Vecchiarelli; A Papi; G Biasc