Apoptotic effect in the glioma cells induced by specific protein extracted from Okinawa Habu (Trimeresurus flavoviridis) venom in relation to oxidative stress
Apoptotic effect in the glioma cells induced by specific protein extracted from Okinawa Habu (Trimeresurus flavoviridis) venom in relation to oxidative stress
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DOI：
10.1016/s0887-2333(03)00010-9
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发表时间：
2003-04-01
影响因子：
3.2
通讯作者：
Morine, N
中科院分区：
文献类型：
--
作者：
Sun, LK;Yoshii, Y;Morine, N
冲绳哈布（Trimeresurus flavoviridis）蛇毒是一种毒性很强的蛇毒，从该蛇毒中提取了一种特异性蛋白质--冲绳哈布脱辅基蛋白-1（OHAP-1）。本研究的目的是探讨OHAP-1是否能诱导胶质瘤细胞凋亡，并探讨其可能的作用机制。测试了三种恶性胶质瘤细胞系。恶性胶质瘤细胞系为大鼠C6和人RBR 17 T、U251。OHAP-1抑制所有细胞系的生长。采用DNA凝胶电泳、DNA流式细胞术和TUNEL法检测细胞凋亡情况。OHAP-1处理后，C6、RBR 17 T和U251细胞中DNA片段化，亚二倍体DNA含量百分比增加，TUNEL阳性细胞。此外，OHAP-1显示L-氨基酸氧化酶（LAAO）活性。为探讨OHAP-1诱导细胞凋亡的机制，采用流式细胞术检测细胞内活性氧（ROS）的变化，免疫组化检测p53蛋白的表达。OHAP-1可诱导神经胶质瘤细胞产生活性氧，并增加p53蛋白的表达。当加入过氧化氢酶或还原型谷胱甘肽（GSH）抗氧化剂时，OHAP-1对三种受试细胞的抑制作用被逆转，其凋亡作用相应地减弱。本研究证实了OHAP-1对某些恶性胶质瘤细胞的凋亡作用，其作用可能是通过促进胶质瘤细胞内ROS的产生和p53蛋白的表达而介导的。提示OHAP-1有望成为临床肿瘤治疗的候选基因。(C)2003爱思唯尔科技有限公司版权所有。
Okinawa Habu (Trimeresurus flavoviridis) venom is well known for its toxic efficacy, from which one kind of specific protein, Okinawa Habu apoxin protein-1 (OHAP-1) has been extracted. The purpose of this study was to investigate whether OHAP-1 could induce apoptosis in some glioma cells, and if so, to elucidate the possible mechanism involved. Three malignant glioma cell lines were tested. The malignant glioma cell lines were rat C6 and human RBR 17T, U251. OHAP-1 inhibited growth of all cell lines. Whether or not the apoptosis had been induced was determined by using DNA gel electrophoresis, DNA flow cytometry and TUNEL assay. After OHAP-1 treatment, DNA fragmentation, an increase in the percentage of subdiploid DNA content, and TUNEL positive cells were found in the C6, RBR17T, and U251 cells. Furthermore, OHAP-1 showed L-amino acid oxidase (LAAO) activity. In order to study the mechanism of apoptosis induced by OHAP-1, the changes of intracellular reactive oxygen species (ROS) were measured using flow cytometry, and the expression of p53 protein was examined using immunohistochemistry. OHAP-1 was found to generate ROS and increase the expression of p53 protein in glioma cells. The inhibiting effect of OHAP-1 on three tested cells was reversed when an antioxidant of either catalase or reduced glutathione (GSH) was added; its apoptotic effect correspondingly became weaker. In this study, the apoptotic effect of OHAP-1 on some malignant glioma cells was confirmed, and it could be that this effect might be mediated through promoting the generation of intracellular ROS and p53 protein expression in glioma cells. It was suggested that OHAP-1 is promising as a potential candidate for clinical tumor therapy. (C) 2003 Elsevier Science Ltd. All rights reserved.