Multicellular tumor spheroid model to evaluate spatio-temporal dynamics effect of chemotherapeutics: application to the gemcitabine/CHK1 inhibitor combination in pancreatic cancer

• Published in: BMC cancer Vol. 12; no. 1; p. 15
• Main Authors: Dufau, Isabelle, Frongia, Céline, Sicard, Flavie, Dedieu, Laure, Cordelier, Pierre, Ausseil, Frédéric, Ducommun, Bernard, Valette, Annie
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 13-01-2012; BioMed Central; BMC
• Subjects: Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis - drug effects; Cancer; Cancer therapies; Carcinoma, Pancreatic Ductal - drug therapy; Carcinoma, Pancreatic Ductal - pathology; Cell cycle; Cell Proliferation - drug effects; CHK1 inhibitor; Combination; Cytotoxicity; Deoxycytidine - analogs & derivatives; Deoxycytidine - pharmacology; DNA damage; DNA Damage - drug effects; DNA repair; Drug resistance; Drug Resistance, Neoplasm - physiology; Evaluation; Gemcitabine; Humans; Kinases; Life Sciences; Models, Biological; Pancreas; Pancreatic cancer; Pancreatic Neoplasms - drug therapy; Pancreatic Neoplasms - pathology; Protein Kinase Inhibitors - pharmacology; Quinolines - pharmacology; Quinuclidines - pharmacology; Spheroids, Cellular - drug effects; Tumor Cells, Cultured; Tumor Microenvironment; Tumor spheroid; Tumors
• ISSN: 1471-2407; 1471-2407
• DOI: 10.1186/1471-2407-12-15

The multicellular tumor spheroid (MCTS) is an in vitro model associating malignant-cell microenvironment and 3D organization as currently observed in avascular tumors.
In order to evaluate the relevance of this model for pre-clinical studies of drug combinations, we analyzed the effect of gemcitabine alone and in combination with the CHIR-124 CHK1 inhibitor in a Capan-2 pancreatic cell MCTS model.
Compared to monolayer cultures, Capan-2 MCTS exhibited resistance to gemcitabine cytotoxic effect. This resistance was amplified in EGF-deprived quiescent spheroid suggesting that quiescent cells are playing a role in gemcitabine multicellular resistance. After a prolonged incubation with gemcitabine, DNA damages and massive apoptosis were observed throughout the spheroid while cell cycle arrest was restricted to the outer cell layer, indicating that gemcitabine-induced apoptosis is directly correlated to DNA damages. The combination of gemcitabine and CHIR-124 in this MCTS model, enhanced the sensitivity to the gemcitabine antiproliferative effect in correlation with an increase in DNA damage and apoptosis.
These results demonstrate that our pancreatic MCTS model, suitable for both screening and imaging analysis, is a valuable advanced tool for evaluating the spatio-temporal effect of drugs and drug combinations in a chemoresistant and microenvironment-depending tumor model.