文章基本信息

标题：Induction of Apoptosis by Disturbing Mitochondrial-Membrane Potential and Cleaving PARP in Jurkat T Cells through Treatment with Acetoxyscirpenol Mycotoxins
本地全文：下载
作者：Dong Hee Lee ; Taesun Park ; Ha Won Kim 等
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：2006
卷号：29
期号：4
页码：648-654
DOI：10.1248/bpb.29.648
出版社：The Pharmaceutical Society of Japan
摘要：Paecilomyces tenuipes is a famous Chinese medicinal entomopathogenic fungus that grows within the larvae of silkworms. 4β-acetoxyscirpendiol (4-MAS), a cytotoxic compound belonging to the scirpenol subfamily of trichothecene mycotoxin, was isolated from Paecilomyces tenuipes . To further elucidate the cytotoxic mechanism of 4-MAS, evidences of its induction of apoptosis, together with the structurally related acetoxyscirpenol moiety mycotoxins (ASMs) such as, 15-acetoxyscirpenol (15-MAS), 4,15-diacetoxyscirpenol (4,15-DAS), and 3α-acetyldiacetoxyscirpenol (TAS), in the human Jurkat T cell line were reported herein. In the MTT reduction and time-course cytotoxicity assays for monitoring cell viability, all the four ASMs that were tested exhibited cytotoxicity; single acetoxylation at C-4 of the scirpenol family resulted in relatively weak cytotoxicity, while acetoxylation at C-15 resulted in strong cytotoxicity regardless of the other acetoxylations at the C-3 and/or C-4 positions. Phosphatidylserine externalization was induced by all the ASMs that were treated at an early phase in a time-dependent manner, showing a typical apoptotic phenomenon, not a necrotic one. The ASMs also reduced the mitochondria's inner-membrane potential (ΔΨm) through flow cytometry analysis after staining these with DiOC₆, a mitochondria-specific and voltage-dependent dye. Acetoxylation of ASM at C-15 increased ΔΨm disruption, but that at C-3 reduced the ΔΨm. The ASMs that were tested also cleaved 113 kDa PARP to an 89-kDa fragment through Western blot assay, suggesting the activation of caspase-3 and/or caspase-7 in the Jurkat T cell. DNA fragmentation was also observed to have been increased in a time-dependent manner by the ASMs that were tested in Jurkat T cells, resulting in the DNA fragmentation intensity order of 4,15-DAS>15-MAS>TAS>4-MAS. These data indicate that the Jurkat T cells that were treated with ASMs underwent typical cascades of apoptotic cell death.
关键词：acetoxyscirpenol;Paecilomyces tenuipes;apoptosis;mitochondrial membrane potential;caspase;DNA fragmentation