LAT1 overexpression and function compensates downregulation of ASCT2 in an in vitro model of renal proximal tubule cell ageing.
Amino acid transporters provide cells neutral amino acids indispensable for growth and proliferation-dependent protein synthesis. This study evaluates whether prolonged serial cell passaging during 6 months (over 50 passages) may induce changes in amino acid transporters properties in Opossum kidney (OK) proximal tubular cells. High passage OK cells exhibit polyploidy, but no difference in the proliferation potential was observed when compared to low passage OK cells. Increased time in culture was accompanied by an increased total, membrane and cytosol protein content. The Na(+)-insensitive [(14)C]-L-leucine uptake was promoted almost exclusively thought LAT1 (~ 90 vs 80%, high versus low passage OK cells). The increased LAT1 protein abundance in high passage OK cells correlated positively with enhanced ability to take up [(14)C]-L-leucine, despite a 4.3-fold decrease in affinity for the substrate. The Na(+)-sensitive [(14)C]-L-alanine transport was decreased by 2.5-fold in high passage OK cells. However, no differences in ASCT2 expression were observed between high and low passage OK cells. It is concluded that OK cells show functional differences in both L-leucine and L-alanine uptake as a function of passage time in culture. The increased expression and activity of LAT1 in high passage OK cells may correspond to a mechanism enabling the cell to develop the hypertrophy response to prolonged cell passaging, when the function of ASCT2 is markedly depressed.
Faculty of Medicine, Institute of Pharmacology and Therapeutics, Porto, Portugal.
Maria João
MJ
Pinho
José Miguel
JM
Cabral
Elisabete
E
Silva
Maria Paula
MP
Serrão
Patrício
P
Soares-da-Silva
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000409:Alanine / Q000378:metabolism; D026962:Amino Acid Transport System ASC / Q000235:genetics*; D000598:Amino Acids, Cyclic / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D016922:Cellular Senescence / Q000235:genetics*; D015536:Down-Regulation; D027282:Fusion Regulatory Protein 1, Heavy Chain / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000378:metabolism; D007700:Kinetics; D027283:Large Neutral Amino Acid-Transporter 1 / Q000096:biosynthesis*; D007930:Leucine / Q000378:metabolism; D009893:Opossums; D011123:Polyploidy; D013347:Subcellular Fractions / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D026962:Amino Acid Transport System ASC; D000598:Amino Acids, Cyclic; D027282:Fusion Regulatory Protein 1, Heavy Chain; D027283:Large Neutral Amino Acid-Transporter 1; C006504:2-aminobicyclo(2,2,1)heptane-2-carboxylic acid; D007930:Leucine; D000409:Alanine