Current evidence suggests that carbonic anhydrase II (CA II) is produced by pancreatic duct cells but not by pancreatic acinar or islet cells. The aim of this study was to determine whether CA II homologous RNA and CA II immunoreactive protein are produced by cell lines established from human pancreatic adenocarcinomas. A 1.7-Kb CA II homologous RNA was detected in poly(A+) RNA isolated from normal human pancreas, normal human liver, and to varying degrees in the cell lines examined. The CA II immunoreactivity corresponding to approximately 30 kD (consistent with the established molecular mass of CA II) was also detected by immunoblotting in normal human pancreas, normal human liver, and some of the cell lines. We also found that the levels of CA II homologous RNA increase in the pancreatic adenocarcinoma cell lines following treatment with the differentiating agent, retinoic acid.