Anti-inflammatory drugs alter amino acid transport in HTC cells.
Following the addition of indomethacin to exponentially growing rat hepatoma cell (HTC) cultures, cells accumulated in the G1 phase. Over the course of several hours, specific changes in membrane transport accompanied this inhibition. Indomethacin stimulated the uptake of 2-aminobicyclo(2,2,1)heptane-2-carboxylic acid (BCH), inhibited the Na+-dependent active transport of alpha-aminoisobutyric acid (AIB), and methylaminoisobutyric acid (MeAIB), but had no effect on the uptake of thymidine, uridine, or 2-deoxyglucose. The stimulation of BCH transport was immediate and reversible. The uptake of BCH was Na+-independent and only slightly depressed by metabolic inhibitors (10 to 20%). Inhibition of AIB and MeAIB uptake, which was of gradual onset, and was observed with several anti-inflammatory drugs and was dependent on the concentration of drugs. Upon removal of drug, AIB and MeAIB transport gradually increased and reached a maximum prior to resumption of DNA synthesis and cell division. Thee effects involved changes in Vmax only. Neither the apparent affinity (Km) for amino acids nor rate of exodus (k, 0.7h-1 for BCH, 1.2h-1 for AIB) were altered. The uptake of MeAIB (and AIB) occurred by a low and a high affinity (Km = 0.27 mM) component. Indomethacin inhibited specifically the high affinity component which was shown to be Na+- and energy-dependent. Although this component was inhibited by treatment with agents that lowered ATP levels or blocked protein synthesis, the anti-inflammatory drugs appeared not to act through these mechanisms.
D000596:Amino Acids / Q000378:metabolism*; D000621:Aminoisobutyric Acids / Q000378:metabolism; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D003513:Cycloheximide / Q000494:pharmacology; D004273:DNA, Neoplasm / Q000096:biosynthesis; D003847:Deoxyglucose / Q000378:metabolism; D007213:Indomethacin / Q000494:pharmacology*; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000096:biosynthesis; D012334:RNA, Neoplasm / Q000096:biosynthesis; D051381:Rats; D013936:Thymidine / Q000378:metabolism; D014529:Uridine / Q000378:metabolism
D000596:Amino Acids; D000621:Aminoisobutyric Acids; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm; D003513:Cycloheximide; D003847:Deoxyglucose; D013936:Thymidine; D014529:Uridine; D007213:Indomethacin