Inflammasomes are intracellular protein complexes that form in response to infections and tissue damage, they play a key role in cancer-related inflammation. This study investigated the expression of inflammasome-related genes, cytokine secretion and caspase-1 enzyme activity in three human pancreatic cancer cell lines: AsPC-1, BxPC-3 and CFPAC-1. As well as co-localization of NLRC4 which is a protein that plays a key role in inflammasome formation and ASC which is an adaptor protein with a crucial role in inflammasome formation. Quantitative PCR showed distinct gene expression patterns of ASC, IL18, IL1β, caspase-1, caspase-8 and NLRP3 across the three cell lines. ASC and IL1β gene expression was significantly different across cell lines. BxPC-3 showed significantly higher IL1β and ASC gene expression compared to AsPC-1 and CFPAC-1. Caspase-1 was overexpressed in BxPC- 3 and was significantly different between BxPC-3 and AsPC-1. IL18 and NLRP3 were highly expressed in AsPC-1 while caspase-8 levels remained similar across the three cell lines. However, NLRC4 gene expression was too low to be included in the statistical analysis. Immunofluorescence demonstrated colocalization of NLRC4 and ASC in AsPC-1 and CFPAC- 1 but not in BxPC-3 suggesting differential inflammasome complex formation. ELISA data showed IL1β protein levels within the normal range of a healthy adult. Caspase-1 assay showed no significant differences in media samples; however, intracellular caspase-1 activity varied significantly, with a notable difference between AsPC-1 and BxPC-3. These findings highlight different inflammasome regulation in pancreatic cancer cell lines with BxPC-3 exhibiting an activated inflammasome profile marked by elevated ASC, IL1β and Caspase-1 expression and secretion which can be attributed to its origin from a primary tumour. In contrast, AsPC-1 and CFPAC-1 showed limited or differential inflammasome assembly potentially influenced by tumor origin and microenvironmental factors. Understanding these molecular distinctions provides insights into the role of inflammasomes in pancreatic cancer.