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Mechanism of nitric oxide‐induced apoptosis in human neuroblastoma SH‐SY5Y cells
Author(s) -
Moriya Ryuichi,
Uehara Takashi,
Nomura Yasuyuki
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)02167-0
Subject(s) - apoptosis , caspase , dna fragmentation , microbiology and biotechnology , caspase 3 , sh sy5y , chemistry , caspase 8 , caspase 9 , mitochondrial permeability transition pore , nitric oxide , mitochondrion , inner mitochondrial membrane , programmed cell death , biology , biochemistry , cell culture , neuroblastoma , genetics , organic chemistry
We have attempted to elucidate the precise mechanism of nitric oxide (NO)‐induced apoptotic neuronal cell death. Enzymatic cleavages of DEVD‐AFC, VDVAD‐AFC, and LEHD‐AFC (specific substrates for caspase‐3‐like protease (caspase‐3 and ‐7), caspase‐2, and caspase‐9, respectively) were observed by treatment with NO. Western blot analysis showed that pro‐forms of caspase‐2, ‐3, ‐6, and ‐7 are decreased during apoptosis. Interestingly, Ac‐DEVD‐CHO, a caspase‐3‐like protease inhibitor, blocked not only the decreases in caspase‐2 and ‐7, but also the formation of p17 from p20 in caspase‐3 induced by NO, suggesting that caspase‐3 exists upstream of caspase‐2 and ‐7. Bongkrekic acid, a potent inhibitor of mitochondrial permeability transition, specifically blocked both the loss of mitochondrial membrane potential and subsequent DNA fragmentation in response to NO. Thus, NO results in neuronal apoptosis through the sequential loss of mitochondrial membrane potential, caspase activation, and degradation of inhibitor of caspase‐activated DNase (CAD) (CAD activation).