Apical uptake of organic cations by human intestinal Caco-2 cells: putative involvement of ASF transporters

The aim of this work was to characterise the intestinal absorption of organic cations, by testing the possibility of involvement of known members of the amphiphilic solute facilitator (ASF) family in this process. For that purpose, the characteristics of the uptake of 1-methyl-4-phenylpyridinium, a model organic cation, at the brush-border membrane of Caco-2 cells were compared with those of the extraneuronal monoamine transporter (EMT)mediated transport. Uptake of [H-3]MPP+ by Caco-2 and 293(hEMT) cells showed pH-dependence: it was significantly reduced (to 86% and 62% of control, respectively) when the pH of the extracellular medium was decreased to 6.2, and increased (to 116% and 136% of control, respectively) when the extracellular pH was increased to 8.2. Uptake of [H-3]MPP+ by Caco-2 cells and 293(hEMT) cells showed potential-dependence: substitution of KCl for NaCl in the incubation medium resulted in a reduction in the inward transport of [H-3]MPP+ (to 70% and 40% of control, respectively). Uptake of [H-3]MPP+ by Caco-2 and 293(hEMT) cells showed only little dependence on Na+: substitution of NaCl of the incubation media with LiCl resulted in a small decrease (of 19% and 14%, respectively) in [H-3]MPP+ uptake. However, when NaCl was substituted with choline chloride, a significant reduction in [H-3]MPP+ uptake by Caco-2 and 293 cells (of 56% and 68%, respectively) was observed. The effect of various compounds on initial rates of [H-3]MPP+ uptake into Caco-2 and 293(hEMT) cells was tested. All compounds tested interacted with the specific [H-3]MPP+ uptake in both cell lines. There was no correlation between the IC(50)s in relation to inhibition of [H-3]MPP+ uptake into Caco-2 cells and into 293(hEMT) cells. Reverse transcriptase-polymerase chain reaction indicates that mRNA of hEMT and of the human organic cation transporter 1 (hOCT1) are present in Caco-2 cells. In conclusion, our results suggest that uptake of organic cations at the brush-border membrane of Caco-2 cells may occur through two distinct Na+-independent transporters belonging to the ASF family: hEMT and hOCT1.