Characterization of primary pancreatic cancer cell lines from iKras*p53* mice.

Abstract

<p>(<b>A</b>) Primary cell line iKras*p53*-1 exhibits epithelial morphology and demonstrates doxycycline dependent Kras* expression (* p<0.05), and pERK1/2 levels at passage 5. (<b>B</b>) The same cell line at passage 12; Kras* expression is still dependent on doxy (*** p<0.001), but pERK1/2 levels do not depend on Kras* expression. (<b>C</b>) A second primary cell line, iKras*p53*-2, has mesenchymal morphology. At passage 6, Kras* expression is dependent on doxy (** p<0.01), and pERK1/2 levels depend on Kras* expression. (<b>D</b>) Analysis at passage 11: Kras* is still regulated by doxy (* p<0.05, ** p<0.01), but pERK1/2 levels remain elevated. (<b>E</b>) Western blot analysis of apoptosis, indicated by cleaved caspase-3 (CC3), and proliferation, measured by proliferating cell nuclear antigen (PCNA), in both iKras*p53*-1 and iKras*p53*-2 cell lines. (<b>F</b>) Immunofluorescence of apoptosis, indicated by cleaved caspase-3 (CC3), in iKras*p53*-2 cells either in the presence of (+48 h) or absence (−48 h) of doxy in the media. DAPI staining marks the nuclei. Scale bar 100 um. (<b>G</b>) Ras pull-down assay demonstrates that Ras activity levels are comparable between iKras*p53* cell lines and cells from KPC tumors.</p

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