Department of Genetics and Cell Biology, Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, China.
Acta Pharmacol Sin. 2011 Dec;32(12):1543-8. doi: 10.1038/aps.2011.130. Epub 2011 Oct 10.
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.
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.
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.
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.
有丝分裂驱动蛋白 Eg5 在双极纺锤体组装中发挥关键作用，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。本研究旨在研究特异性 Eg5 抑制剂二甲苯并嗪对胰腺癌细胞迁移和侵袭的影响。
使用人胰腺癌细胞系 PANC1、EPP85、BxPC3、CFPAC1 和 AsPAC1。使用免疫荧光显微镜检查 Eg5 表达。通过划痕愈合和 Transwell 测定分析细胞迁移和侵袭。使用磺酰罗丹明 B 和 MTT 测定法检查细胞增殖。通过分子建模研究分析二甲苯并嗪与 Eg5 的结合，并使用 MANT-ADP 试剂检查 ADP 释放率。
在 5 种胰腺癌细胞系中，Eg5 表达上调了 9-16 倍。用二甲苯并嗪（3 和 10 μmol/L）处理 PANC1 胰腺癌细胞 24 h 可浓度依赖性地抑制癌细胞的迁移能力。癌细胞的侵袭能力也降低了。然而，用二甲苯并嗪（3 和 10 μmol/L）处理 PANC1 细胞 24 h 对其增殖没有可检测的影响，而当癌细胞用药物处理 72 h 时，癌细胞的增殖受到抑制。分子建模研究表明，二甲苯并嗪可以通过降低 ADP 释放率来变构抑制 Eg5 的马达结构域 ATP 酶。
二甲苯并嗪抑制 PANC1 胰腺癌细胞的迁移和侵袭，而不抑制细胞增殖。这些发现为针对胰腺癌化疗的 Eg5 靶向提供了新的见解。
