Enhancement of the Antiproliferative Activity of Gemcitabine by Modulation of c-Met Pathway in Pancreatic Cancer

Author(s): Amir Avan, Karl Quint, Francesco Nicolini, Niccola Funel, Adam E. Frampton, Mina Maftouh, Serena Pelliccioni, Gerrit J. Schuurhuis, Godefridus J. Peters, Elisa Giovannetti.

Journal Name: Current Pharmaceutical Design

Volume 19 , Issue 5 , 2013

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.

Keywords: Pancreatic ductal adenocarcinoma, c-Met pathway, cancer stem cells, crizotinib, gemcitabine, Capan-1-gemcitabine-resistant cells (Capan-1-R), sulforhodamine-B-assay, flow cytometry, combination-index method, cell-growth

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Article Details

VOLUME: 19
ISSUE: 5
Year: 2013
Page: [940 - 950]
Pages: 11
DOI: 10.2174/1381612811306050940
Price: $58

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