Expression levels of a kinesin-13 microtubule depolymerase modulates the effectiveness of anti-microtubule agents.
Expression levels of a kinesin-13 microtubule depolymerase modulates the effectiveness of anti-microtubule agents.
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DOI：
10.1371/journal.pone.0011381
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发表时间：
2010-06-30
期刊：
影响因子：
3.7
通讯作者：
Rogers SL
中科院分区：
文献类型：
--
作者：
Schimizzi GV;Currie JD;Rogers SL
化疗药物通常靶向微管细胞骨架作为破坏癌细胞有丝分裂和增殖的手段。抗微管药物抑制微管动力学，从而在分裂细胞激活有丝分裂检查点时触发凋亡。微管动力学是由微管相关蛋白（MAP）调节的，然而，我们缺乏对抗微管药物如何与MAP功能性相互作用的全面了解。在这份报告中，我们测试的假设，即细胞水平的微管解聚酶，在这种情况下驱动蛋白-13，调节微管破坏药物秋水仙碱的有效性。我们在果蝇S2细胞中使用了RNA干扰（RNAi），高通量显微镜和延时视频显微镜的组合来识别特定的MAP，驱动蛋白样蛋白10A（KLP 10A），这有助于抗微管药物秋水仙碱的功效。KLP 10A是整个细胞周期中必需的微管解聚酶。我们发现，在S2细胞中KLP 10A的耗竭赋予抵抗秋水仙素诱导的微管解聚到更大程度上比其他几个不稳定的MAP的耗竭。使用基于图像的测定，我们确定当用2 μM秋水仙素处理1小时后，对照细胞保留58%（±2%SEM）的微管聚合物，而通过RNAi去除KLP 10A的细胞保留74%（±1%SEM）。同样，KLP 10A-GFP的过表达导致对秋水仙素引起的微管解聚的敏感性增加。我们的研究结果表明，由药理学试剂的微管不稳定的功效是依赖于微管解聚酶的细胞表达。这些发现表明，Kif 2A（人类驱动蛋白-13家族成员）的表达水平可能是评估抗微管化疗有效性的有吸引力的生物标志物。了解MAP表达水平如何影响抗微管药物的作用，可能有助于评估癌症治疗的可能模式。
Chemotheraputic drugs often target the microtubule cytoskeleton as a means to disrupt cancer cell mitosis and proliferation. Anti-microtubule drugs inhibit microtubule dynamics, thereby triggering apoptosis when dividing cells activate the mitotic checkpoint. Microtubule dynamics are regulated by microtubule-associated proteins (MAPs); however, we lack a comprehensive understanding about how anti-microtubule agents functionally interact with MAPs. In this report, we test the hypothesis that the cellular levels of microtubule depolymerases, in this case kinesin-13 s, modulate the effectiveness of the microtubule disrupting drug colchicine. We used a combination of RNA interference (RNAi), high-throughput microscopy, and time-lapse video microscopy in Drosophila S2 cells to identify a specific MAP, kinesin-like protein 10A (KLP10A), that contributes to the efficacy of the anti-microtubule drug colchicine. KLP10A is an essential microtubule depolymerase throughout the cell cycle. We find that depletion of KLP10A in S2 cells confers resistance to colchicine-induced microtubule depolymerization to a much greater extent than depletion of several other destabilizing MAPs. Using image-based assays, we determined that control cells retained 58% (±2%SEM) of microtubule polymer when after treatment with 2 µM colchicine for 1 hour, while cells depleted of KLP10A by RNAi retained 74% (±1%SEM). Likewise, overexpression of KLP10A-GFP results in increased susceptibility to microtubule depolymerization by colchicine. Our results demonstrate that the efficacy of microtubule destabilization by a pharmacological agent is dependent upon the cellular expression of a microtubule depolymerase. These findings suggest that expression levels of Kif2A, the human kinesin-13 family member, may be an attractive biomarker to assess the effectiveness of anti-microtubule chemotherapies. Knowledge of how MAP expression levels affect the action of anti-microtubule drugs may prove useful for evaluating possible modes of cancer treatment.
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影响因子：
14.8
作者：
Rogers, Stephen L.;Rogers, Gregory C.
通讯作者：
Rogers, Gregory C.
DOI：
10.1083/jcb.200812052
发表时间：
2009-04-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Mulder AM;Glavis-Bloom A;Moores CA;Wagenbach M;Carragher B;Wordeman L;Milligan RA
通讯作者：
Milligan RA
DOI：
10.1073/pnas.90.24.11598
发表时间：
1993-12-15
影响因子：
11.1
作者：
UPPULURI, S;KNIPLING, L;WOLFF, J
通讯作者：
WOLFF, J
影响因子：
4.3
作者：
Hedrick, David G.;Stout, Jane R.;Walczak, Claire E.
通讯作者：
Walczak, Claire E.
影响因子：
64.5
作者：
Ogawa, T;Nitta, R;Hirokawa, N
通讯作者：
Hirokawa, N