BACKGROUND Malignant cells require increased amounts of amino acids, in particular glutamine and leucine, to support DNA and protein biosynthesis. Although plasma concentrations in the center of solid tumors can be much lower than normal circulating levels, it is still unknown how tumor cells can survive despite low amino acid levels. We examined the effects of glutamine or leucine deprivation on cell growth and amino acid transport activity in two human hepatoma cell lines, SK-Hep and HepG2. METHODS We studied the transport of glutamine, leucine, alanine, and arginine. The carrier-mediated uptake of 3H-amino acids was determined in cells cultured in normal and amino acid-deprived media. RESULTS The growth of both cell lines was dependent on the concentration of glutamine and leucine. In SK-Hep, there was a significant increase in initial rate glutamine transport activity in the glutamine-deprived group, attributable to an increase in transporter affinity (Km; 0.6 mmol/L [control], 385 +/- 43 mumol/L; 0.1 mmol/L, 221 +/- 11 mumol/L; P < 0.01). At low glutamine concentration, the saturable Na(+)-independent uptake of leucine and arginine as well as the Na(+)-dependent uptake of alanine increased significantly in both SK-Hep and HepG2. Similarly, in leucine-deprived SK-Hep cells, leucine uptake increased twofold, but the change was attributable to an enhanced transporter capacity (Vmax; 0.2 mmol/L [control], 38,900 +/- 700; 0.0 mmol/L, 75,900 +/- 4,900 pmol/mg protein per minute; P < 0.001). CONCLUSIONS Adaptive increases in initial rate amino acid transport activities were elicited by glutamine and leucine deprivation in these two human hepatoma cell lines. Decreased extracellular amino acid levels encountered by tumors in vivo may elicit similar adaptive responses that contribute to the maintenance of cytoplasmic levels of amino acids essential for growth.