Induction of apoptosis and ubiquitin hydrolase gene expression by human serum factors in the early phase of acute myocardial infarction.
The physiologic events leading to apoptosis in myocardial infarction and the molecules involved in the death process have not been clarified unequivocally. We developed a method to search for serum factors that induce apoptosis of human cells, using serum obtained from patients within 1 day of the onset of acute myocardial infarction (AMI). Serum factors were found to have the ability to increase the caspase-3 activity levels in human RSa cells, which are susceptible to apoptosis inducers. The factors obtained from AMI patients by elution at about 0.5 mol/L KCl from a dye-ligand column were named AMI-SFs (serum factors from AMI). Electrophoretic analysis showed DNA fragmentation in AMI-SF-treated RSa cells, but not in RSa cells treated with fractions from AMI patients 1 week after clinical onset of illness. AMI-SF-induced DNA fragmentation was also demonstrated by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling analysis, whereas a suppression of fragmentation was seen in RSa cells treated with AMI-SFs in combination with a caspase-3 inhibitor. The increase in caspase-3 activity was not inhibited by neutralizing antibodies to tumor necrosis factor-alpha, interleukin-6, human interferon-beta, or interferon-gamma. Polymerase chain reaction-based messenger RNA differential display and Northern blotting revealed an increase in the messenger RNA expression level of human ubiquitin hydrolase in AMI-SF-treated RSa cells. Antisense oligonucleotides for ubiquitin hydrolase inhibited the increase in caspase-3 activity. These findings suggested that serum from AMI patients in the acute phase contains factors that induce apoptosis, possibly by inducing the expression of the ubiquitin hydrolase gene, at least in the human cells tested.
Hasegawa
Ritsuko
R
Department of Medicine and Clinical Oncology, Graduate School of Medicine, Chiba University, Chiba City, Japan.
Kita
Kazuko
K
Hasegawa
Rei
R
Fusejima
Kenji
K
Fukuzawa
Shigeru
S
Wano
Chieko
C
Watanabe
Shigeru
S
Saisho
Hiromitsu
H
Masuda
Yoshiaki
Y
Nomura
Fumio
F
Suzuki
Nobuo
N
D000328:Adult; D000368:Aged; D017209:Apoptosis / Q000187:drug effects; D001685:Biological Factors / Q000097:blood* / Q000494:pharmacology; D053148:Caspase 3; D020169:Caspases / Q000096:biosynthesis / Q000235:genetics; D002461:Cell Line, Transformed; D005591:Chemical Fractionation; D053938:DNA Fragmentation; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D020287:In Situ Nick-End Labeling; D008297:Male; D008875:Middle Aged; D009203:Myocardial Infarction / Q000097:blood* / Q000473:pathology; D012333:RNA, Messenger / Q000378:metabolism; D013869:Thiolester Hydrolases / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors; D043222:Ubiquitin Thiolesterase
D001685:Biological Factors; D012333:RNA, Messenger; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases