RRC ID 12174
Author Kondo T, Matsuda T, Kitano T, Takahashi A, Tashima M, Ishikura H, Umehara H, Domae N, Uchiyama T, Okazaki T.
Title Role of c-jun expression increased by heat shock- and ceramide-activated caspase-3 in HL-60 cell apoptosis. Possible involvement of ceramide in heat shock-induced apoptosis.
Journal J Biol Chem
Abstract Ceramide has emerged as a lipid mediator in apoptosis induced by a variety of stresses. As we previously showed that the activation of AP-1, a nuclear transcription factor was indispensable to ceramide-induced apoptosis in human leukemia HL-60 cells (Sawai, H., Okazaki, T., Yamamoto, H., Okano, H., Takeda, Y., Tashima, M., Sawada, H., Okuma, M., Ishikura, H., Umehara, H., and Domae, N. (1995) J. Biol. Chem. 270, 27326-27331), the role and mechanism of heat shock (HS)-increased c-jun expression in apoptosis was here investigated. HS increased morphological changes compatible with apoptosis in human leukemia HL-60 cells, and induced ceramide generation and sphingomyelin hydrolysis with an increase of neutral magnesium-dependent sphingomyelinase activity. When HS failed to induce apoptosis in HS-resistant HL-60 cells, ceramide generation was not detected, suggesting that ceramide was involved in downstream signals required for HS-induced apoptosis. Both HS and N-acetylsphingosine (C(2)-ceramide) increased the expression of c-jun/c-fos mRNAs with the peak 2 h after treatment. When we examined whether the inhibition of c-jun expression by its antisense oligodeoxynucleotides (AS) blocked HS- or C(2)-ceramide-induced apoptosis, AS of c-jun gene inhibited apoptotic morphological changes and DNA fragmentation whereas did not sense oligodeoxynucleotides. Moreover, a synthetic tetrapeptide, acetyl-Asp-Met-Gln-Asp-aldehyde (DMQD-CHO), which inhibited the formation of active form of caspase-3 more efficiently than those of caspase-4, -6, -7, and -8, blocked both caspase-3 like activity, c-jun expression and apoptosis induced by HS or C(2)-ceramide, although DMQD-CHO did not affect HS-induced ceramide generation. These results suggested that the ceramide was generated through sphingomyelin hydrolysis by HS-activated neutral, magnesium-dependent sphingomyelinase and that subsequent c-jun expression through activation of caspase-3 played a role in HS-induced HL-60 cell apoptosis.
Volume 275(11)
Pages 7668-76
Published 2000-3-17
DOI 10.1074/jbc.275.11.7668
PMID 10713077
MeSH Apoptosis* Caspase 3 Caspase Inhibitors Caspases / metabolism* Cell Division Ceramides / metabolism* Enzyme Activation HL-60 Cells Heat-Shock Response / physiology* Humans Leukemia, Promyelocytic, Acute / metabolism* Leukemia, Promyelocytic, Acute / therapy Oligodeoxyribonucleotides, Antisense / pharmacology Oligopeptides / pharmacology Proto-Oncogene Proteins c-fos / genetics Proto-Oncogene Proteins c-fos / metabolism Proto-Oncogene Proteins c-jun / genetics Proto-Oncogene Proteins c-jun / metabolism* RNA, Messenger / biosynthesis RNA, Neoplasm / biosynthesis Signal Transduction Sphingomyelin Phosphodiesterase / metabolism Transcription Factor AP-1 / metabolism
IF 4.106
Times Cited 71
DNA material pHSP90 Human hsp 90 alpha cDNA (RDB01127)