L-2,5-dihydrophenylalanine, an inducer of cathepsin-dependent apoptosis in human promyelocytic leukemia cells (HL-60).
L-2,5-Dihydrophenylalanine (DHPA), a phenylalanine analogue, induced apoptosis in human promyelocytic leukemia cells (HL-60). This apoptosis was demonstrated by morphological changes of the cells, such as fragmentation of nuclei and chromatin condensation, and by some evidence found in biochemical analysis, such as DNA ladder and activation of caspase 3. The DHPA-induced apoptosis was prevented by a pan-caspase inhibitor, Z-VAD-fmk, and a cysteine protease inhibitor, E-64d, which inhibits calpains and cathepsin B and L. A calpain inhibitor, Z-LL-H, did not affect this apoptosis. A cathepsin B specific inhibitor, CA074-Me, prevented only chromatin condensation. However, E-64d and a cathepsin L specific inhibitor, Z-FY(t-Bu)-dmk, protected the cells from both chromatin condensation and oligonucleosomal DNA fragmentation. As proceeding to the apoptotic process, the activities of both cathepsin B and L increased gradually. These results indicated that DHPA was an inducer of cathepsin-dependent apoptosis in HL-60 cells.
Kiso
T
T
Graduate School of Science, Osaka City University, Osaka, Japan.
Usuki
Y
Y
Ping
X
X
Fujita
K
K
Taniguchi
M
M
D017209:Apoptosis / Q000187:drug effects*; D002154:Calpain / Q000037:antagonists & inhibitors; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002403:Cathepsins / Q000378:metabolism / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000187:drug effects; D053138:Cyclohexenes; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D053938:DNA Fragmentation; D005456:Fluorescent Dyes; D018922:HL-60 Cells; D006801:Humans; D009707:Nucleosomes / Q000187:drug effects / Q000378:metabolism; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology
D061945:Caspase Inhibitors; D002843:Chromatin; D053138:Cyclohexenes; D015853:Cysteine Proteinase Inhibitors; D005456:Fluorescent Dyes; D009707:Nucleosomes; D011480:Protease Inhibitors; C004174:2,5-dihydrophenylalanine; D010649:Phenylalanine; D002403:Cathepsins; C505407:CASP3 protein, human; D002154:Calpain; D053148:Caspase 3; D020169:Caspases