Description
Pancreatic ductal adenocarcinoma (PDAC) cells display significant metabolic flexibility and can utilize fatty acid metabolism to support tumor growth and survival. While fatty acid oxidation (FAO) is primarily known for its role in energy production, disruption of this pathway may also alter intracellular lipid homeostasis. In this study, we investigated the role of fatty acid β-oxidation in regulating lipid storage in pancreatic cancer cells by silencing ACADS, a key mitochondrial enzyme involved in short-chain fatty acid oxidation. Transfection efficiency was first confirmed using Cy3-conjugated scrambled siRNA, which demonstrated greater than 90% transfection efficiency. Successful knockdown of ACADS was further validated by Western blot analysis, showing more than 70% reduction in ACADS protein levels following siRNA treatment. Inhibition of fatty acid β-oxidation impaired the ability of pancreatic cancer cells to metabolize fatty acids, resulting in intracellular lipid accumulation. BODIPY neutral lipid staining revealed a marked increase in lipid droplets following ACADS knockdown. To further confirm lipid droplet formation, we examined the expression of perilipin, a lipid droplet–associated protein that regulates lipid storage and protects triacylglycerol droplets from lipolysis. Western blot analysis demonstrated increased perilipin expression in ACADS-silenced cells, further supporting enhanced lipid storage. Interestingly, we also observed a decrease in mitochondrial oxidative phosphorylation proteins following ACADS inhibition, suggesting broader metabolic consequences associated with impaired fatty acid utilization.
Together, these findings indicate that disruption of fatty acid β-oxidation promotes lipid droplet accumulation and alters lipid homeostasis in pancreatic cancer cells. These results highlight the importance of fatty acid metabolism in regulating intracellular lipid balance and suggest that targeting lipid metabolic pathways may represent a potential strategy for pancreatic cancer therapy.