%0 Journal Article
%T Role of c-MET Inhibitors in Overcoming Drug Resistance in Spheroid Models of Primary Human Pancreatic Cancer and Stellate Cells
%A Amir Avan
%A Btissame El Hassouni
%A Elisa Giovannetti
%A Giulia Mantini
%A Godefridus J. Peters
%A Ilaria Carnevale
%A Luciano Saso
%A Mark Buijs
%A Matthias L£¿hr
%A Niccola Funel
%A Omidreza Firuzi
%A Pei Pei Che
%A Rainer Heuchel
%A Stefano Coppola
%A Thomas Schmidt
%J Archive of "Cancers".
%D 2019
%R 10.3390/cancers11050638
%X Pancreatic stellate cells (PSCs) are a key component of tumor microenvironment in pancreatic ductal adenocarcinoma (PDAC) and contribute to drug resistance. c-MET receptor tyrosine kinase activation plays an important role in tumorigenesis in different cancers including PDAC. In this study, effects of PSC conditioned medium (PCM) on c-MET phosphorylation (by immunocytochemistry enzyme-linked immunosorbent assay (ELISA)) and drug response (by sulforhodamine B assay) were investigated in five primary PDAC cells. In novel 3D-spheroid co-cultures of cyan fluorescence protein (CFP)-firefly luciferase (Fluc)-expressing primary human PDAC cells and green fluorescence protein (GFP)-expressing immortalized PSCs, PDAC cell growth and chemosensitivity were examined by luciferase assay, while spheroids¡¯ architecture was evaluated by confocal microscopy. The highest phospho-c-MET expression was detected in PDAC5 and its subclone sorted for ¡°stage specific embryonic antigen-4¡± (PDAC5 (SSEA4)). PCM of cells pre-incubated with PDAC conditioned medium, containing increased hepatocyte growth factor (HGF) levels, made PDAC cells significantly more resistant to gemcitabine, but not to c-MET inhibitors. Hetero-spheroids containing both PSCs and PDAC5 (SSEA4) cells were more resistant to gemcitabine compared to PDAC5 (SSEA4) homo-spheroids. However, c-MET inhibitors (tivantinib, PHA-665752 and crizotinib) were equally effective in both spheroid models. Experiments with primary human PSCs confirmed the main findings. In conclusion, we developed spheroid models to evaluate PSC¨CPDAC reciprocal interaction, unraveling c-MET inhibition as an important therapeutic option against drug resistant PDAC
%K pancreatic cancer
%K three-dimensional culture
%K drug resistance
%K cancer-associated fibroblasts
%K primary cultures
%U https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6562408/