Rotenone inhibits mammalian cell proliferation by inhibiting microtubule assembly through tubulin binding
Rotenone inhibits mammalian cell proliferation by inhibiting microtubule assembly through tubulin binding
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DOI：
10.1111/j.1742-4658.2007.06004.x
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发表时间：
2007-09-01
期刊：
影响因子：
5.4
通讯作者：
Panda, Dulal
中科院分区：
文献类型：
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作者：
Srivastava, Pallavi;Panda, Dulal
鱼藤酮是一种广泛使用的杀虫剂，已被证明能抑制哺乳动物细胞的增殖并解聚细胞微管。在本研究中，鱼藤酮对微管组装的影响与其抑制细胞增殖和有丝分裂的能力有关。我们发现鱼藤酮抑制HeLa和MCF-7细胞的增殖，其半数最大抑制浓度分别为0.2+/-0.1mU和0.4+/-0.1mU/M。在其有效抑制浓度范围内，鱼藤酮解聚了两种细胞类型的纺锤体微管。然而，与HeLa细胞相比，它对MCF-7细胞间期微管的影响要强得多。鱼藤酮抑制活的HeLa细胞中微管的重组，表明它可以抑制微管的生长速度。此外，它在较低的有效浓度范围内缩短了HeLa细胞的中心体间距，在相对较高的浓度范围内诱导了多极纺锤体的形成。它还增加了动粒区域检查点蛋白BubR1的水平。鱼藤酮在体外对微管组装和GTP水解率均有抑制作用。还可抑制秋水仙碱与微管蛋白的结合，扰乱微管蛋白的二级结构，降低微管蛋白的内源性色氨酸荧光和微管蛋白-1-苯氨基萘-8-磺酸复合体的外源荧光，提示其与微管蛋白结合。微管蛋白-鱼藤酮复合体的解离常数为3+/-0.6µM。目前的数据表明，鱼藤酮通过干扰微管组装动力学来阻止有丝分裂和抑制细胞增殖。
Rotenone, a widely used insecticide, has been shown to inhibit mammalian cell proliferation and to depolymerize cellular microtubules. In the present study, the effects of rotenone on the assembly of microtubules in relation to its ability to inhibit cell proliferation and mitosis were analyzed. We found that rotenone inhibited the proliferation of HeLa and MCF-7 cells with half maximal inhibitory concentrations of 0.2 +/- 0.1 mu M and 0.4 +/- 0.1 mu M, respectively. At its effective inhibitory concentration range, rotenone depolymerized spindle microtubules of both cell types. However, it had a much stronger effect on the interphase microtubules of MCF-7 cells compared to that of the HeLa cells. Rotenone suppressed the reassembly of microtubules in living HeLa cells, suggesting that it can suppress microtubule growth rates. Furthermore, it reduced the intercentrosomal distance in HeLa cells at its lower effective concentration range and induced multipolar-spindle formation at a relatively higher concentration range. It also increased the level of checkpoint protein BubR1 at the kinetochore region. Rotenone inhibited both the assembly and the GTP hydrolysis rate of microtubules in vitro. It also inhibited the binding of colchicine to tubulin, perturbed the secondary structure of tubulin, and reduced the intrinsic tryptophan fluorescence of tubulin and the extrinsic fluorescence of tubulin-1-anilinonaphthalene-8-sulfonic acid complex, suggesting that it binds to tubulin. A dissociation constant of 3 +/- 0.6 mu M was estimated for tubulin-rotenone complex. The data presented suggest that rotenone blocks mitosis and inhibits cell proliferation by perturbing microtubule assembly dynamics.