Butyrate and the other short chain fatty acids (SCFAs) are the most abundant anions in the colonic lumen. Also, butyrate is the preferred energy source for colonocytes and has been shown to regulate colonic electrolyte and fluid absorption. Previous studies from our group have demonstrated that HCO₃^-/SCFA^- anion exchange process is one of the major mechanisms of butyrate transport across the purified human colonic apical membrane vesicles and the apical membrane of human colonic adenocarcinoma cell line, Caco2, and suggested that it is mainly mediated via monocarboxylate transporter MCT-1 isoform. However, little is known regarding the regulation of SCFA transport by various hormones and signal transduction pathways. Therefore, the current studies were undertaken to examine whether hydrocortisone and phorbol ester, PMA are involved in a possible regulation of butyrate/anion exchange process in Caco2 cells. Butyrate/anion exchange process was assessed by measuring a pH-driven ¹⁴C-butyrate uptake in Caco2 cells. Our results demonstrated that 24 h incubation with PMA (1 µM) significantly increased ¹⁴C-butyrate uptake compared to incubation with 4PMA (inactive form). In contrast, incubation with hydrocortisone had no significant effect on butyrate uptake in Caco2 cells compared to vehicle (ethanol) alone. The induction of butyrate uptake by PMA appeared to be via an increase in the V_max of the transport process with no significant changes in the K_m of the transporter for butyrate. Parallel to the increase in the V_max of ¹⁴C-butyrate uptake, MCT-1 protein level was also increased in response to PMA incubation. Our studies demonstrated that the butyrate/anion exchange was increased in response to PMA treatment along with the induction in the level of MCT1 expression in Caco2 cells.