Dimethylenastron suppresses human pancreatic cancer cell migration and invasion in vitro via allosteric inhibition of mitotic kinesin Eg5
Dimethylenastron suppresses human pancreatic cancer cell migration and invasion in vitro via allosteric inhibition of mitotic kinesin Eg5
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Dimethylenastron 通过有丝分裂驱动蛋白 Eg5 的变构抑制在体外抑制人胰腺癌细胞迁移和侵袭
DOI：
10.1038/aps.2011.130
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发表时间：
2011-12-01
影响因子：
8.2
通讯作者：
Zhou, Jun
中科院分区：
文献类型：
--
作者：
Sun, Xiao-dong;Shi, Xing-juan;Zhou, Jun
目的：有丝分裂运动蛋白Eg5在双极纺锤体组装中起着至关重要的作用，其抑制剂在临床前研究中显示出令人印象深刻的抗癌活性。本研究旨在探讨Eg5特异性抑制剂二甲基萘醌对胰腺癌细胞迁移和侵袭的影响。方法：采用人胰腺癌细胞系PANC1、EPP85、BxPC3、CFPAC1、AsPAC1。免疫荧光显微镜检测Eg5的表达。用伤口愈合和transwell实验分析细胞迁移和侵袭。采用硫磺胺B和MTT法检测细胞增殖。采用分子模拟方法分析二甲基萘醌与Eg5的结合，并采用MANT-ADP试剂检测ADP的释放速率。结果：5种胰腺癌细胞系中Eg5表达上调9 ~ 16倍。3、10 μmol/L二甲基萘醌作用胰腺胰腺细胞24 h后，癌细胞的迁移能力呈浓度依赖性抑制。治疗也降低了癌细胞的侵袭能力。3 μmol/L和10 μmol/L二甲基萘醌作用于胰腺胰腺1细胞24 h后对其增殖无明显影响，作用72 h后癌细胞增殖受到抑制。分子模型研究表明，二甲基萘醌可通过降低ADP的释放速率来变构抑制Eg5的运动域atp酶。结论：二甲萘醌可抑制胰腺癌细胞PANC1的迁移和侵袭，但不抑制细胞增殖。这些发现为胰腺癌化疗靶向Eg5的机制提供了新的见解。
Aim:The mitotic kinesin Eg5 plays a critical role in bipolar spindle assembly, and its inhibitors have shown impressive anticancer activity in preclinical studies. This study was undertaken to investigate the effect of dimethylenastron, a specific inhibitor of Eg5, on the migration and invasion of pancreatic cancer cells.Methods:Human pancreatic cancer cell lines PANC1, EPP85, BxPC3, CFPAC1, and AsPAC1 were used. Eg5 expression was examined using immunofluorescence microscopy. Cell migration and invasion were analyzed with wound healing and transwell assays. Cell proliferation was examined using sulforhodamine B and MTT assays. The binding of dimethylenastron to Eg5 was analyzed with a molecular modeling study, and the ADP release rate was examined with the MANT-ADP reagent.Results:Eg5 expression was 9–16-fold up-regulated in the 5 pancreatic cancer cell lines. Treatment of PANC1 pancreatic cancer cells with dimethylenastron (3 and 10 μmol/L) for 24 h suppressed the migratory ability of the cancer cells in a concentration-dependent manner. The invasion ability of the cancer cells was also reduced by the treatment. However, treatment of PANC1 cells with dimethylenastron (3 and 10 μmol/L) for 24 h had no detectable effect on their proliferation, which was inhibited when the cancer cells were treated with the drug for 72 h. Molecular modeling study showed that dimethylenastron could allosterically inhibit the motor domain ATPase of Eg5 by decreasing the rate of ADP release.Conclusion:Dimethylenastron inhibits the migration and invasion of PANC1 pancreatic cancer cells, independent of suppressing the cell proliferation. The findings provide a novel insight into the mechanisms of targeting Eg5 for pancreatic cancer chemotherapy.