Antimitotic effect of the retinoid 4-oxo-fenretinide through inhibition of tubulin polymerization: a novel mechanism of retinoid growth-inhibitory activity

Mol Cancer Ther. 2009 Dec;8(12):3360-8. doi: 10.1158/1535-7163.MCT-09-0798.

Abstract

The retinoid 4-oxo-N-(4-hydroxyphenyl)retinamide (4-oxo-4-HPR), a metabolite of fenretinide (4-HPR) present in plasma of 4-HPR-treated patients, is very effective in inducing growth inhibition and apoptosis in several cancer cell lines. 4-Oxo-4-HPR and 4-HPR have different mechanisms of action because 4-oxo-4-HPR, unlike 4-HPR, causes marked cell accumulation in G2-M phase. Here, we investigated the molecular events involving 4-oxo-4-HPR-induced cell cycle perturbation in ovarian (A2780 and IGROV-1) and breast (T47D, estrogen receptor+ and BT-20, estrogen receptor-) cancer cells. 4-Oxo-4-HPR induced a delay of mitosis (with mitotic index increasing 5- to 6-fold in all cell lines) without progression beyond the anaphase, as shown by cyclin B1 expression. 4-Oxo-4-HPR induced multipolar spindle formation and phosphorylation of BUBR1, resulting in activation of the spindle checkpoint. Multipolar spindles were not due to impairment of pole-focusing process, loss of centrosome integrity, or modulation of the expression levels of molecules associated with spindle aberrations (Kif 1C, Kif 2A, Eg5, Tara, tankyrase-1, centractin, and TOGp). We show here that 4-oxo-4-HPR targets microtubules because, in treated cells, it interfered with the reassembly of cold-depolymerized spindle microtubules and decreased the polymerized tubulin fraction. In cell-free assays, 4-oxo-4-HPR inhibited tubulin polymerization (50% inhibition of microtubule assembly at 5.9 micromol/L), suggesting a direct molecular interaction with tubulin. In conclusion, by showing that 4-oxo-4-HPR causes mitotic arrest through antimicrotubule activities, we delineate a new molecular mechanism for a retinoid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Antimitotic Agents / pharmacology*
  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology
  • Cell Cycle / drug effects
  • Cell Line, Tumor
  • Female
  • Fenretinide / analogs & derivatives*
  • Fenretinide / pharmacology
  • Gene Expression Regulation, Neoplastic / drug effects
  • Growth Inhibitors / pharmacology
  • Humans
  • Kinesins / genetics
  • Microfilament Proteins / genetics
  • Microscopy, Fluorescence
  • Microtubule-Associated Proteins / genetics
  • Microtubules / drug effects
  • Microtubules / metabolism
  • Mitotic Index
  • Ovarian Neoplasms / genetics
  • Ovarian Neoplasms / metabolism
  • Ovarian Neoplasms / pathology
  • Polymers / metabolism
  • Retinoids / pharmacology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Spindle Apparatus / drug effects
  • Tankyrases / genetics
  • Tubulin / metabolism*

Substances

  • 4-oxofenretinide
  • Actins
  • Antimitotic Agents
  • CKAP5 protein, human
  • Growth Inhibitors
  • KIF11 protein, human
  • KIF1C protein, human
  • KIF2A protein, human
  • Microfilament Proteins
  • Microtubule-Associated Proteins
  • Polymers
  • Retinoids
  • TRIOBP protein, human
  • Tubulin
  • centractin
  • Fenretinide
  • Tankyrases
  • TNKS protein, human
  • Kinesins