Heat-induced apoptosis via caspase-3 activation in tumour cells carrying mutant p53.
Apoptosis plays an important role in heat-induced cell death. However, the mechanism of heat-induced apoptosis has not yet been elucidated. In the present study, the signal transduction pathway underlying heat-induced apoptosis was investigated in heat-resistant HeLa cells carrying mutant p53 gene and heat sensitive HeLa cells that had been transduced with an antisense TNF gene. Induction of mutant p53, but not p21/WAF-1, was observed after heat treatment of both the resistant and sensitive cells. Heat-induced cytotoxicity was not inhibited in either cells with interleukin-1beta-converting enzyme (ICE: caspase-1) like protease inhibitor Ac-YVAD-CHO. In contrast, there was 48% and 63% inhibition of cytotoxicity in HeLa and transfectants, respectively, with a caspase-3 inhibitor (Ac-DEVD-CHO). Heat-induced apoptosis was also prevented by administration of Ac-DEVD-CHO in both cells. In addition, an augmentation of heat-induced cytotoxicity in transfectants was almost completely inhibited by Ac-DEVD-CHO. Further, caspase-3 mRNA expression was increased remarkably in heat-treated HeLa cells and transfectants. Taken together, these results suggest that activation of caspase-3 is involved in the signal transduction pathway of heat-induced apoptosis of the tumour cells carrying mutant p53.
Kobayashi
D
D
Department of Laboratory Diagnosis, Sapporo Medical University, School of Medicine, Japan.
Watanabe
N
N
Yamauchi
N
N
Okamoto
T
T
Tsuji
N
N
Sasaki
H
H
Sato
T
T
Niitsu
Y
Y
D001483:Base Sequence; D053148:Caspase 3; D020169:Caspases / Q000235:genetics / Q000378:metabolism*; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000378:metabolism; D017931:DNA Primers; D004789:Enzyme Activation; D015971:Gene Expression Regulation, Enzymologic; D016158:Genes, p53; D050884:HSP72 Heat-Shock Proteins; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000037:antagonists & inhibitors; D006358:Hot Temperature; D006801:Humans; D009154:Mutation; D009842:Oligopeptides / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors
C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D017931:DNA Primers; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D009842:Oligopeptides; D012333:RNA, Messenger; C096429:L 709049; D013482:Superoxide Dismutase; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases