Active roles of caspase-3 in human gastric carcinoma cell death by apoptosis inducing nucleosides from CD57+HLA-DRbright natural suppressor cell line.
The six apoptosis inducing nucleosides (AINs), which were isolated by high performance liquid chromatography from CD57+HLA-DRbright natural suppressor (57.DR-NS) cell cultures, induced apoptosis in human gastric carcinoma (GCIY) cells demonstrating the accumulation of sub-G1 DNA content and morphological changes. By means of DNA unwinding assay, it has been revealed that DNA strand breaks were initially involved in the apoptotic cell death of GCIY cells treated with AINs followed by activation of caspase cascades, especially caspase-3. Actually, the cleavage of fluorogenic substrate for caspase-3 was identified in the reaction. Whereas, the addition of caspase-3 inhibitor into the reaction prevented the cleavage of fluorogenic substrate for caspase-3 and resulted in the blockage of the sub-G1 DNA accumulation and DNA fragmentation as apoptotic signals. Thus, it was definitely elucidated that the activation of caspase-3 displayed a key feature during AIN-induced apoptosis in GCIY cells.
Li
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Department of Immunology, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-0071, Japan.
Guo
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M
Mori
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Mori
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T
D017209:Apoptosis / Q000187:drug effects; D051033:Apoptosis Inducing Factor; D018959:CD57 Antigens / Q000502:physiology; D053148:Caspase 3; D061945:Caspase Inhibitors; D020169:Caspases / Q000502:physiology*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D004273:DNA, Neoplasm / Q000032:analysis; D004789:Enzyme Activation; D005420:Flavoproteins / Q000302:isolation & purification / Q000494:pharmacology*; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000502:physiology; D006801:Humans; D015151:Immunoblotting; D008565:Membrane Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D018613:Microscopy, Confocal; D013274:Stomach Neoplasms / Q000201:enzymology / Q000473:pathology*; D050378:T-Lymphocytes, Regulatory / Q000737:chemistry* / Q000276:immunology; D014407:Tumor Cells, Cultured
C494829:AIFM1 protein, human; D051033:Apoptosis Inducing Factor; D018959:CD57 Antigens; D061945:Caspase Inhibitors; D004273:DNA, Neoplasm; D005420:Flavoproteins; D006684:HLA-DR Antigens; D008565:Membrane Proteins; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases