DELINEATING THE INTRACELLULAR AND EXTRACELLULAR ROLE OF MATRIX METALLOPROTEINASE 14 IN PRIMARY AND METASTATIC PANCREATIC CANCERS BxPC-3 AND HPAF-II

Using flow cytometry and two forms of hypoxic induction, chemical (CoCl2) and gaseous (Tri-gas chamber), this study highlighted the advantages of quantitative analyses of MMP-14 expression (intracellularly/extracellularly) for one primary (BxPC-3) and one metastatic (HPAF-II) pancreatic cancer line. No significant changes in MMP-14 expression were observed for pancreatic cancer lines using CoCl2. Increased expression levels of MMP-14 were observed using tri-gas incubation maintaining oxygen levels at 2% intracellularly for HPAF-II, but not BxPC-3. A combination of 2D/3D cell culturing techniques were also used to examine changes in cellular behavior/morphology after hypoxic exposure and a 24-hour reoxygenation cycle. BxPC-3 cells showed a greater propensity toward oxidative damage caused by reoxygenation using 2D culturing techniques. Using 3D biomimetic culturing techniques, reoxygenated BxPC-3 cells did not undergo significant apoptosis or necrosis. These results suggest that changes in cellular metabolism and behavior depend on both phenotype and culture scaffolding.