zVAD-fmk, unlike BocD-fmk, does not inhibit caspase-6 acting on 14-3-3/Bad pathway in apoptosis of p815 mastocytoma cells.
In a preliminary study, we found that benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (zVAD- fmk), unlike Boc-aspartyl(OMe)-fluoromethylketone (BocD-fmk), at usual dosage could not prevent genistein-induced apoptosis of p815 mastocytoma cells. This study was undertaken to reveal the mechanism underlying the incapability of zVAD-fmk in preventing this type of apoptosis. We observed that 14-3-3 protein level was reduced in genistein-treated cells and that BocD-fmk but not zVAD-fmk prevented the reduction of 14-3-3 protein level and the release of Bad from 14-3-3. We also demonstrated that truncated Bad to Bcl-xL interaction in genistein- treated cells was prevented by BocD-fmk but not by zVAD-fmk treatment. Our data indicate that BocD- fmk, compared to zVAD-fmk, has a certain preference for inhibiting 14-3-3/Bad signalling pathway. We also elucidated that this differential efficacy of BocD-fmk and zVAD-fmk resulted from the different effect in inhibiting caspase-6 and that co-treatment of zVAD-fmk and caspase-6 specific inhibitor substantially prevented genistein-induced apoptosis. Our data shows that caspase-6 plays a role on Bad/14-3-3 pathway in genistein-induced apoptosis of p815 cells, and that the usual dose of zVAD-fmk, in contrast to BocD-fmk, did not prevent caspase-6 acting on 14-3-3/Bad-mediated event.
Yee
Su-Bog
SB
Department of Anatomy and Cell Biology (BK21 program), Dong-A University College of Medicine and Medical, Science Research Center, Busan 602-714, Korea.
Baek
Soo Jin
SJ
Park
Hwan Tae
HT
Jeong
Seung Hun
SH
Jeong
Jin Hee
JH
Kim
Tae Hyun
TH
Kim
Jong-Min
JM
Jeong
Byung Kap
BK
Park
Bong Soo
BS
Kwon
Taeg Kyu
TK
Yoon
Il
I
Yoo
Young Hyun
YH
D048948:14-3-3 Proteins / Q000378:metabolism*; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001593:Benzyl Compounds / Q000494:pharmacology*; D053178:Caspase 6 / Q000378:metabolism*; D061945:Caspase Inhibitors; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019833:Genistein / Q000494:pharmacology; D006845:Hydrocarbons, Fluorinated / Q000494:pharmacology*; D034801:Mastocytoma; D051379:Mice; D008928:Mitochondria / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D051025:bcl-Associated Death Protein / Q000378:metabolism*
D048948:14-3-3 Proteins; D000590:Amino Acid Chloromethyl Ketones; D001593:Benzyl Compounds; C476029:Boc-D-FMK; D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D006845:Hydrocarbons, Fluorinated; D051025:bcl-Associated Death Protein; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D019833:Genistein; D053178:Caspase 6