title: Enhancement of the antiproliferative activity of gemcitabine by modulation of c-Met pathway in pancreatic cancer.
abstract: Pancreatic-ductal-adenocarcinoma (PDAC) is amongst the most lethal malignancies, mainly because of its metastatic spread and multifactorial chemoresistance. Since c-Met is a marker of pancreatic-cancer-stem-cells (CSC), playing a key role in metastasis and chemoresistance, this study evaluated the therapeutic potential of the novel c-Met/ALK inhibitor crizotinib against PDAC cells, including the Capan-1-gemcitabine-resistant cells (Capan-1-R). Crizotinib inhibited PDAC cell-growth with IC50 of 1.5 μM in Capan-1-R, and synergistically enhanced the antiproliferative and proapoptotic activity of gemcitabine, as detected by sulforhodamine-B-assay, flow cytometry and combination-index method. Capan-1-R had higher expression of the CSC markers CD44+/CD133+/CD326+, but their combined expression was significantly reduced by crizotinib, as detected by quantitative-RT-PCR and FACS-analysis. Similarly, Capan-1-R cells had significantly higher protein-expression of c-Met (≈2-fold), and increased migratory activity, which was reduced by crizotinib (e.g., > 50% reduction of cell-migration in Capan-1-R after 8-hour exposure, compared to untreated-cells), in association with reduced vimentin expression. Capan-1-R had also significantly higher mRNA expression of the gemcitabine catabolism-enzyme CDA, potentially explaining the higher CDA activity and statistically significant lower levels of gemcitabine-nucleotides in Capan-1-R compared to Capan-1, as detected by Liquid-chromatography-massspectrometry. Conversely, crizotinib significantly reduced CDA expression in both Capan-1 and Capan-1-R cells. In aggregate, these data show the ability of crizotinib to specifically target CSC-like-subpopulations, interfere with cell-proliferation, induce apoptosis, reduce migration and synergistically interact with gemcitabine, supporting further studies on this novel therapeutic approach for PDAC.
authors.affiliation: Dept. Medical Oncology, VUmc Cancer Center Amsterdam, VU University Medical Center, CCA room 1.52, De Boelelaan 1117, 1081HV Amsterdam, The Netherlands.
authors.forename: Amir
authors.initials: A
authors.lastname: Avan
authors.forename: Karl
authors.initials: K
authors.lastname: Quint
authors.forename: Francesco
authors.initials: F
authors.lastname: Nicolini
authors.forename: Niccola
authors.initials: N
authors.lastname: Funel
authors.forename: Adam E
authors.initials: AE
authors.lastname: Frampton
authors.forename: Mina
authors.initials: M
authors.lastname: Maftouh
authors.forename: Serena
authors.initials: S
authors.lastname: Pelliccioni
authors.forename: Gerrit J
authors.initials: GJ
authors.lastname: Schuurhuis
authors.forename: Godefridus J
authors.initials: GJ
authors.lastname: Peters
authors.forename: Elisa
authors.initials: E
authors.lastname: Giovannetti
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation / Q000187:drug effects; D002853:Chromatography, Liquid / Q000379:methods; D000077547:Crizotinib; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm; D004357:Drug Synergism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020128:Inhibitory Concentration 50; D013058:Mass Spectrometry / Q000379:methods; D014411:Neoplastic Stem Cells / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D047428:Protein Kinase Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism; D011720:Pyrazoles / Q000008:administration & dosage / Q000494:pharmacology*; D011725:Pyridines / Q000008:administration & dosage / Q000494:pharmacology*; D000093542:Gemcitabine
chemical_list: D000964:Antimetabolites, Antineoplastic; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011725:Pyridines; D003841:Deoxycytidine; D000077547:Crizotinib; D019859:Proto-Oncogene Proteins c-met; D000093542:Gemcitabine