Growth inhibitory effect of L-canavanine against MIA PaCa-2 pancreatic cancer cells is not due to conversion to its toxic metabolite canaline.
L-Canavanine (CAV) is an arginine (ARG) analog isolated from the jack bean, Canavalia ensiformis. CAV becomes incorporated into cellular proteins of MIA PaCa-2 human pancreatic cancer cells and the aberrant, canavanyl proteins are not preferentially degraded. Hydrolytic cleavage of CAV to canaline (CAN) and urea is mediated by arginase. CAN is a potent metabolite that inactivates vitamin B6-containing enzymes and may inhibit cell growth. To determine the presence of arginase and its specificity for ARG and CAV in MIA PaCa-2 cells, a radiometric assay, which quantifies the 14C released from the hydrolytic cleavage of L-[guanidino-14C]ARG or L-[guanidinooxy-14C]CAV mediated by arginase, was employed. Insignificant amounts of 14CO2 were released when cells were exposed to [14C]CAV or to [14C]ARG. Pancreatic cancer cells secrete a negligible amount of arginase. Cytotoxic effects of CAN and CAV were compared on cells exposed to varying concentrations of ARG. These studies provide evidence that CAV's cytotoxic effects on MIA PaCa-2 cells cannot be attributed to conversion to the active metabolite CAN. A slower and decreased hydrolysis of CAV by arginase allows CAV to persist and increases its chances of incorporating into proteins in these cells. Lack of considerable amounts of arginase in the pancreas makes CAV a worthy candidate for further studies in pancreatic cancer.
Swaffar
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Division of Basic Pharmaceutical Sciences, School of Pharmacy, Northeast Louisiana University, Monroe 71209, USA. pyswaffar@alpha.nlu.edu
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D000613:Aminobutyrates / Q000378:metabolism* / Q000633:toxicity; D001119:Arginase / Q000378:metabolism; D002172:Canavanine / Q000378:metabolism / Q000494:pharmacology*; D002245:Carbon Dioxide / Q000032:analysis / Q000378:metabolism; D002250:Carbon Radioisotopes / Q000032:analysis; D002455:Cell Division / Q000187:drug effects; D004396:Coloring Agents / Q000737:chemistry; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D011874:Radiometry / Q000379:methods; D013379:Substrate Specificity; D013778:Tetrazolium Salts / Q000737:chemistry; D013844:Thiazoles / Q000737:chemistry; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000473:pathology
D000613:Aminobutyrates; D002250:Carbon Radioisotopes; D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D002245:Carbon Dioxide; D002172:Canavanine; C001926:canaline; D001119:Arginase; C022616:thiazolyl blue