1. Development of bioluminescent chick chorioallantoic membrane (CAM) models for primary pancreatic cancer cells: a platform for drug testing.
- Author
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Rovithi M, Avan A, Funel N, Leon LG, Gomez VE, Wurdinger T, Griffioen AW, Verheul HM, and Giovannetti E
- Subjects
- Adenocarcinoma drug therapy, Adenocarcinoma genetics, Adenocarcinoma pathology, Animals, Carcinogenesis pathology, Carcinoma, Pancreatic Ductal drug therapy, Carcinoma, Pancreatic Ductal genetics, Carcinoma, Pancreatic Ductal pathology, Cell Proliferation drug effects, Cell Proliferation genetics, Chickens, Crizotinib pharmacology, Crizotinib therapeutic use, DNA Mutational Analysis, Deoxycytidine analogs & derivatives, Deoxycytidine pharmacology, Deoxycytidine therapeutic use, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, Hepatocyte Nuclear Factor 6 genetics, Hepatocyte Nuclear Factor 6 metabolism, MicroRNAs genetics, MicroRNAs metabolism, Pancreatic Neoplasms drug therapy, Pancreatic Neoplasms genetics, SOX9 Transcription Factor genetics, SOX9 Transcription Factor metabolism, Sequence Analysis, DNA, Tumor Cells, Cultured, Gemcitabine, Pancreatic Neoplasms, Chorioallantoic Membrane metabolism, Drug Evaluation, Preclinical, Luminescent Measurements methods, Models, Biological, Pancreatic Neoplasms pathology
- Abstract
The aim of the present study was to develop chick-embryo chorioallantoic membrane (CAM) bioluminescent tumor models employing low passage cell cultures obtained from primary pancreatic ductal adenocarcinoma (PDAC) cells. Primary PDAC cells transduced with lentivirus expressing Firefly-luciferase (Fluc) were established and inoculated onto the CAM membrane, with >80% engraftment. Fluc signal reliably correlated with tumor growth. Tumor features were evaluated by immunohistochemistry and genetic analyses, including analysis of mutations and mRNA expression of PDAC pivotal genes, as well as microRNA (miRNA) profiling. These studies showed that CAM tumors had histopathological and genetic characteristic comparable to the original tumors. We subsequently tested the modulation of key miRNAs and the activity of gemcitabine and crizotinib on CAM tumors, showing that combination treatment resulted in 63% inhibition of tumor growth as compared to control (p < 0.01). These results were associated with reduced expression of miR-21 and increased expression of miR-155. Our study provides the first evidence that transduced primary PDAC cells can form tumors on the CAM, retaining several histopathological and (epi)genetic characteristics of original tumors. Moreover, our results support the use of these models for drug testing, providing insights on molecular mechanisms underlying antitumor activity of new drugs/combinations.
- Published
- 2017
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