Anti-microtubule activity of tubeimoside I and its colchicine binding site of tubulin.
Anti-microtubule activity of tubeimoside I and its colchicine binding site of tubulin.
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DOI：
10.1007/s00280-007-0635-0
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发表时间：
2008-09
影响因子：
3
通讯作者：
Yu, Tingxi
中科院分区：
文献类型：
--
作者：
Ma, Rundi;Song, Gang;You, Wenbing;Yu, Lijian;Su, Weiming;Liao, Mingneng;Zhang, Yongping;Huang, Laizhen;Zhang, Xiaoyu;Yu, Tingxi
关键词：
Tubeimoside I （TBMS1）是从Bolbostemma paniculatum （Maxim.）块茎中分离得到的。Franquet。TBMS1具有较强的抗肿瘤活性。本研究旨在探讨TBMS1的抗微管作用及其与微管蛋白的结合位点。用MTT法检测TBMS1对细胞生长的抑制作用。采用高效液相色谱法测定了人鼻咽癌CNE-2Z细胞株（CNE-2Z）对TBMS1的摄取动力学。以猪脑为原料，在高摩尔浓度缓冲液中经聚合-解聚两个循环制备微管蛋白（MTP）。通过浊度测定和沉降试验确定TBMS1对MTP聚合的抑制作用；采用免疫荧光显微镜和免疫印迹技术研究TBMS1与CNE-2Z细胞微管蛋白的相互作用。通过竞争性结合实验测试TBMS1抑制已知微管蛋白配体结合的能力。TBMS1对CNE-2Z细胞的生长抑制活性为72 h， IC50值为16.7 μM。HPLC分析显示，CNE-2Z细胞对TBMS1的摄取初始缓慢，在18 h左右逐渐达到最大，TBMS1作用于CNE-2Z细胞（25 μM, 3 h）足以导致微管网络破坏。免疫印迹分析显示，经TBMS1处理的细胞胞质微管蛋白比例呈时间和浓度依赖性增加。TBMS1不抑制长春花碱与微管蛋白的结合。在TBMS1的存在下，秋水仙碱与微管蛋白的结合受到抑制。TBMS1是一种抗微管药物，其微管蛋白结合位点为微管蛋白的秋水仙碱结合位点。
Tubeimoside I (TBMS1) was isolated from the tubers of Bolbostemma paniculatum (Maxim.) Franquet. TBMS1 shows potent anti-tumor activity. The present study was conducted to investigate the anti-microtubule role of TBMS1 and its binding site of tubulin. Cell growth inhibition was measured by MTT after treatment with TBMS1. Uptake kinetics of TBMS1 by human nasopharyngeal carcinoma CNE-2Z cell line (CNE-2Z) was assayed by HPLC. Microtubule protein (MTP) was prepared from porcine brain through two cycles of polymerization–depolymerization in a high molarity buffer. Inhibition of MTP polymerization induced by TBMS1 was determined by a turbidity measurement and a sedimentation assay; the interactions of TBMS1 with tubulin within CNE-2Z cells were investigated by immunofluorescence microscopy and immunoblotting. TBMS1 was tested for its ability to inhibit binding of known tubulin ligands through competitive binding assay. TBMS1 displayed growth inhibitory activity against CNE-2Z cells with IC50 value of 16.7 μM for 72 h. HPLC analysis of TBMS1 uptake by CNE-2Z cells displayed the initial slow TBMS1 uptake and then gradually reaching an maximum uptake near 18 h. CNE-2Z cells treated with TBMS1 (25 μM, 3 h) were sufficient to cause the microtubular network disruption. Immunoblot analysis showed that the proportion of cytosolic tubulin of cells treated with TBMS1 increased in a time- and concentration-dependent manner. TBMS1 did not inhibit the binding of vinblastine to tubulin. Colchicine binding to tubulin was inhibited in the presence of TBMS1. TBMS1 is an anti-microtubule agent, and its binding site of tubulin is the colchicine binding site of tubulin.
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影响因子：
5.8
作者：
BAI, R;FRIEDMAN, SJ;HAMEL, E
通讯作者：
HAMEL, E
影响因子：
7.5
作者：
Mitsui-Saito, M;Ohkubo, S;Nakahata, N
通讯作者：
Nakahata, N
影响因子：
5.8
作者：
BAI, R;PETTIT, GR;HAMEL, E
通讯作者：
HAMEL, E
影响因子：
5.8
作者：
Edler, MC;Fernandez, AM;Barrows, LR
通讯作者：
Barrows, LR
DOI：
10.1073/pnas.91.9.3964
发表时间：
1994-04-26
影响因子：
11.1
作者：
DAMATO, RJ;LIN, CM;HAMEL, E
通讯作者：
HAMEL, E