title: CPUYJ039, a newly synthesized benzimidazole-based compound, is proved to be a novel inducer of apoptosis in HCT116 cells with potent KSP inhibitory activity.
abstract: OBJECTIVE
  This study investigated the antiproliferative and apoptotic activities of CPUYJ039, a newly synthesized benzimidazole-based kinesin spindle protein (KSP) inhibitor, on HCT116 cell lines.
  
  
  METHODS
  KSP-inhibitory activity was tested using the malachite-green method. The in-vitro cell proliferation inhibitory activity of the sample was measured using WST reagent. Flow-cytometric evaluation of cellular DNA content was performed. Apoptotic cells were quantified by annexin V-FITC-PI double staining. To confirm that the cytotoxic activity was a consequence of KSP inhibition, microtubule morphology and DNA segregation were observed by double staining of microtubules and DNA. The expression of Bcl-2 and Bax in CPUYJ039-treated HCT116 cells was detected by Western blotting.
  
  
  RESULTS
  CPUYJ039 was evaluated and proved to have potent inhibitory activities in the KSP ATPase and HCT116 cell proliferation assays. CPUYJ039 inhibited the proliferation of HCT116 cells in a dose- and time-dependent manner and markedly induced G2/M phase cell-cycle arrest with characteristic monoastral spindles and subsequent cell death in HCT116 cells, which was associated with an increase of the Bax/Bcl-2 ratio.
  
  
  CONCLUSIONS
  These results suggest that CPUYJ039 may be a novel inducer of apoptosis in HCT116 cells with potent KSP inhibitory activity.
authors: Jiangsu Key Laboratory of Carcinogenesis and Intervention, Nanjing, China.
authors: Cheng
authors: C
authors: Jiang
authors: Lei
authors: L
authors: Yang
authors: Wu-Tong
authors: WT
authors: Wu
authors: Qing-Long
authors: QL
authors: Guo
authors: Qi-Dong
authors: QD
authors: You
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D050256:Antimitotic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001562:Benzimidazoles / Q000494:pharmacology*; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D045325:HCT116 Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D016547:Kinesins / Q000037:antagonists & inhibitors*; D008870:Microtubules / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013997:Time Factors; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D050256:Antimitotic Agents; C494826:BAX protein, human; D001562:Benzimidazoles; C570061:CPUYJ039; C470243:KIF11 protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D000251:Adenosine Triphosphatases; D016547:Kinesins