title: Dimethylenastron suppresses human pancreatic cancer cell migration and invasion in vitro via allosteric inhibition of mitotic kinesin Eg5.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors: Department of Genetics and Cell Biology, Tianjin Key Laboratory of Protein Science, College of Life Sciences, Nankai University, China.
authors: Xiao-dong
authors: XD
authors: Sun
authors: Xing-juan
authors: XJ
authors: Shi
authors: Xiao-ou
authors: XO
authors: Sun
authors: You-guang
authors: YG
authors: Luo
authors: Xiao-jing
authors: XJ
authors: Wu
authors: Chang-fu
authors: CF
authors: Yao
authors: Hai-yang
authors: HY
authors: Yu
authors: Deng-wen
authors: DW
authors: Li
authors: Min
authors: M
authors: Liu
authors: Jun
authors: J
authors: Zhou
mesh_terms: D000494:Allosteric Regulation; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D006801:Humans; D016547:Kinesins / Q000037:antagonists & inhibitors*; D009361:Neoplasm Invasiveness / Q000517:prevention & control*; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D011799:Quinazolines / Q000494:pharmacology*; D013871:Thiones / Q000494:pharmacology*
chemical_list: C470243:KIF11 protein, human; D011799:Quinazolines; D013871:Thiones; C518969:dimethylenastron; D016547:Kinesins