Knockout studies of Panc1 cells

Abstract: Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal form of cancerwith very few available treatment options of which none has great effect.Cancer cells and stromal cells such as stellate cells which exist in abundancein PDAC interact by crosstalk, resulting in a tumorigenic collective response.With the help of a previously developed 3D co-culture spheroid model theeffect of a CRISPR/cas9 knockout of the cellular communication cetworkfactor 1 (CCN1) gene together with gemcitabine (GEM) treatment has beeninvestigated in terms of Panc1 cell viability and gene expression. Spheroidsconsisting of wild-type and knockout cell lines, each identified by westernblots were cultured, imaged and treated. Viability assays and RNA extractionfollowed by PCR showed that the viability of the cancer cells in the spheroidswere higher for the cells with CCN1 knockout. Cancer cells were also coculturedwith stellate cells with the goal of investigating the effect of thecellular crosstalk on chemoresistance.