The differentiation of colon cancer cell lines is associated with changes in calcium homeostasis. Concomitantly there are changes in the expression of some calcium transporters and G-protein-coupled receptors, which are capable of altering cytosolic-free calcium levels. Recent studies associate alterations in calcium transporter expression with tumourigenesis, such as changes in specific isoforms of the plasma membrane calcium ATPase (PMCA) in breast cancer cell lines. In this study, we examined the expression of PMCA isoforms in the HT-29 colon cancer cell line using two methods of differentiation (sodium butyrate-mediated and spontaneous post-confluency induced differentiation). Our studies show that differentiation of HT-29 colon cancer cells is associated with the up-regulation of the PMCA isoform PMCA4 but no significant alteration in PMCA1. These results suggest that PMCA4 may be important and have a specific role in colon cells as well as being significant in colon cancer tumourigenesis.