Na+-independent phosphate transport in Caco2BBE cells

P-i transport in epithelia has both Na+-dependent and Na+-independent components, but so far only Na+-dependent transporters have been characterized in detail and molecularly identified. Consequently, in the present study, we initiated the characterization and analysis of intestinal Na+-independent P-i transport using an in vitro model, Caco2BBE cells. Only Na+-independent P-i uptake was observed in these cells, and Pi uptake was dramatically increased when cells were incubated in high-P-i DMEM (4 mM) from 1 day to several days. No response to low-P-i medium was observed. The increased P-i transport was mainly caused by V-max changes, and it was prevented by actinomycin D and cycloheximide. P-i transport in cells grown in 1 mM P-i (basal DMEM) decreased at pH > 7.5, and it was inhibited with proton ionophores. P-i transport in cells incubated with 4 mM P-i increased with alkaline pH, suggesting a preference for divalent phosphate. P-i uptake in cells in 1 mM Pi was completely inhibited only by Pi and partially inhibited by phosphonoformate, oxalate, DIDS, SITS, SO42-, HCO3-, and arsenate. This inhibition pattern suggests that more than one P-i transporter is active in cells maintained with 1 mM P-i. Phosphate transport from cells maintained at 4 mM P-i was only partially inhibited by phosphonoformate, oxalate, and arsenate. Attempts to identify the responsible transporters showed that multifunctional anion exchangers of the Slc26 family as well as members of Slc17, Slc20, and Slc37 and the P-i exporter xenotropic and polytropic retrovirus receptor 1 are not involved.