Growth-dependent AIB and meAIB uptake in LLC-PK1 cells: effects of differentiation inducers and of TPA.

Cultured pig kidney cells designated LLC-PK1, previously shown to acquire Na+-dependent concentrative transport of hexoses as the cells become growth arrested, also show Na+-dependent concentrative uptake of the amino acid analogs alpha-aminoisobutyric acid (AIB) and (methyl) meAIB. This A system-like transport is most active in sparse, growing cultures and becomes stepped down at confluence. The cell/medium equilibrium distribution ratio of the lipophilic cation tetraphenylphosphonium ion (TPP+) decreases in parallel fashion, suggesting that a decrease in membrane potential may be a major factor in the stepdown. Differentiation inducers (hexamethylene bisacetamide) and phosphodiesterase inhibitors (theophylline, methylisobutyl xanthine) accelerate the stepdown, but even in the presence of these compounds addition of the tumor promoter 12-0-tetradecanoylphorbol-13-acetate (TPA) results in the maintenance of a high level of AIB and meAIB uptake. In all these respects the changes in A system-like amino acid transport are the reciprocal of those seen for concentrative hexose transport, although the driving force appears to be the same for both systems. The TPA analogs phorbol and 4-0-methyl TPA which are inactive in tumor promotion are inactive in this system as well. In confluent, already stepped-down cultures, addition of TPA leads to a rapid (2-6 hour) stimulation of AIB and meAIB uptake. The enhancement is sensitive to cycloheximide and actinomycin D. The ouabain-sensitive fraction of meAIB uptake is not markedly changed in the TPA-enhanced uptake, nor is the TPP+ distribution ratio elevated in TPA-treated cells, making it unlikely that the TPA effect is through an alteration in the membrane potential.