Cytoprotective effect of arginine deiminase on taxol-induced apoptosis in DU145 human prostate cancer cells.
We purified and partially sequenced a cytostatic protein from the ASC-17D Sertoli cell-conditioned media (rSCCM) showing a molecular weight of 90 kDa with homodimeric composition. N-terminal amino acid analysis revealed that the protein was homologous to the arginine deiminase (ADI) of Mycoplasma arginini. We found ADI enzyme activity in rSCCM and the abolishment of the growth inhibitory effect by the supplement of L-arginine. Thus, we confirmed that the cytostatic activity in rSCCM was due to the depletion of extracellular L-arginine by ADI. Apparent increase of cell death or DNA fragmentation was not observed in DU145 cells cultured in the presence of ADI. Incubation of DU145 cancer cells with taxol resulted in a marked DNA fragmentation, whereas pretreatment with ADI or cycloheximide protected the cells from taxol-induced apoptosis. Preincubation of the cells with ADI inhibited S35-methionine incorporation into protein synthesis in a dose dependent manner. These data suggest that ADI-induced arginine depletion may inhibit protein synthesis, and result in the protection of apoptotic cell death that requires new protein synthesis.
Kang
S W
SW
Department of Pharmacology, College of Medicine, Korea University, Seoul.
Kang
H
H
Park
I S
IS
Choi
S H
SH
Shin
K H
KH
Chun
Y S
YS
Chun
B G
BG
Min
B H
BH
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001120:Arginine / Q000378:metabolism; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D002460:Cell Line; D017077:Culture Media, Conditioned; D003513:Cycloheximide / Q000494:pharmacology; D019610:Cytoprotection; D053938:DNA Fragmentation; D016193:G1 Phase; D006801:Humans; D006867:Hydrolases / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008969:Molecular Sequence Data; D009569:Nitric Oxide / Q000378:metabolism; D017239:Paclitaxel / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D011500:Protein Synthesis Inhibitors / Q000378:metabolism; D051381:Rats; D016415:Sequence Alignment; D012708:Sertoli Cells / Q000378:metabolism; D014407:Tumor Cells, Cultured
D017077:Culture Media, Conditioned; D008070:Lipopolysaccharides; D011500:Protein Synthesis Inhibitors; D009569:Nitric Oxide; D001120:Arginine; D003513:Cycloheximide; D007770:L-Lactate Dehydrogenase; D006867:Hydrolases; C019723:arginine deiminase; D017239:Paclitaxel