Gao Li, Shan Bao-en, Chen Jing, Liu Jian-hui, Song Da-xiang, Zhu Bao-cheng
College of Life Science, Hebei University, Baoding 071002, China.
Acta Pharmacol Sin. 2005 Mar;26(3):369-76. doi: 10.1111/j.1745-7254.2005.00052.x.
To examine the effect of venom from the spider Macrothele raven on cell proliferation and cytotoxicity in human cervical carcinoma, HeLa cells.
Morphological and biochemical signs of apoptosis appeared using acridine orange-ethidium bromide (AO/EB) staining. Marked morphological changes in HeLa cells after treatment with spider venom were observed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Cell proliferation and cytotoxicity were determined by [methyl-3H] thymidine assay ([3H]TdR) and lactate dehydrogenase (LDH) release, respectively. DNA fragmentation and cell cycle distribution were monitored using flow cytometry. In addition, Western blot analysis was used to evaluate the level of caspase-3 expression. In vivo examination of the inhibition of the size of tumors in nude mice treated with spider venom was measured.
Marked morphological changes were observed using AO/EB staining, SEM and TEM assay. Spider venom at concentrations of 10-40 mg/L caused dose- and time-dependent inhibition of HeLa cell proliferation. The ratio of apoptosis and necrosis increased. The activity of caspase-3 was upregulated after spider venom treatment. In vivo study of tumor size revealed that tumors significantly decreased in size from controls to tumors treated for 3 weeks with spider venom (P<0.05).
The inhibition of HeLa cells by the venom of the spider Macrothele raveni was carried out in three ways: induction of apoptosis, necrosis of toxicity damage and direct lysis. Spider venom is a novel anti-tumor material both in vitro and in vivo.
研究雷氏大疣蛛毒液对人宫颈癌HeLa细胞增殖及细胞毒性的影响。
采用吖啶橙-溴化乙锭（AO/EB）染色观察凋亡的形态学和生化特征。利用扫描电子显微镜（SEM）和透射电子显微镜（TEM）观察蜘蛛毒液处理后HeLa细胞明显的形态学变化。分别通过[甲基-³H]胸腺嘧啶核苷测定法（[³H]TdR）和乳酸脱氢酶（LDH）释放来测定细胞增殖和细胞毒性。使用流式细胞术监测DNA片段化和细胞周期分布。此外，采用蛋白质免疫印迹分析评估半胱天冬酶-3的表达水平。测定了经蜘蛛毒液处理的裸鼠体内肿瘤大小的抑制情况。
通过AO/EB染色、SEM和TEM检测观察到明显的形态学变化。浓度为10 - 40 mg/L的蜘蛛毒液对HeLa细胞增殖产生剂量和时间依赖性抑制。凋亡和坏死比例增加。蜘蛛毒液处理后半胱天冬酶-3的活性上调。肿瘤大小的体内研究表明，从对照组到经蜘蛛毒液处理3周的肿瘤，肿瘤大小显著减小（P<0.05）。
雷氏大疣蛛毒液对HeLa细胞的抑制作用通过三种方式实现：诱导凋亡、毒性损伤坏死和直接裂解。蜘蛛毒液在体外和体内均是一种新型抗肿瘤物质。
