Comparative in vitro cytotoxic effects of taxol and its major human metabolite 6 alpha-hydroxytaxol

Cancer Chemother Pharmacol. 1995;36(2):129-35. doi: 10.1007/BF00689197.

Abstract

Taxol is metabolized by the liver microsomal cytochrome P450 enzyme system into its principal metabolite 6 alpha-hydroxytaxol (6HT). In the present in vitro studies 6HT was compared to taxol with respect to its effects on tubulin depolymerization, mitotic arrest, clonogenic survival and apoptosis in HL-60 cells. 6HT was generated by incubating taxol with human liver microsomes in a NADPH-generating system. HL-60 cells were incubated for 24 h with either taxol or 6HT, washed and placed in drug-free suspension or cultured for colony growth in agarose. For the suspension and colony culture growth of the cells, the IC50 concentrations of 6HT were 500 +/- 46 and 350 +/- 37 nM, while those of taxol were 3.2 +/- 0.2 and 2.8 +/- 0.5 nM, respectively. Immediately after a 24-h exposure of HL-60 cells to 50 nM taxol, electrophoresis of genomic DNA from HL-60 cells revealed an internucleosomal DNA fragmentation 'ladder'. In addition, 39% of the cells were arrested in mitosis and 16% showed the morphologic features of apoptosis. In contrast, an identical treatment with 6HT resulted in the mitotic arrest of only 2.8% of the cells, with 4.0% displaying apoptosis (P < 0.01); internucleosomal DNA fragmentation was not observed. 6HT was also significantly less effective than taxol in inhibiting the temperature-induced depolymerization of microtubules in a cell-free system. However, at equipotent concentrations, the effect of 6HT on tubulin depolymerization, mitotic arrest or apoptosis was similar to that of taxol. In addition, at concentrations of taxol or 6HT at or below their IC50, there was little tubulin depolymerization, mitotic arrest or apoptosis. The results presented here show that the biotransformation of taxol to 6HT substantially detoxifies taxol.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Apoptosis / drug effects
  • Cell Division / drug effects
  • Cell Line
  • Cell Survival / drug effects
  • Cell-Free System
  • DNA Damage
  • DNA, Neoplasm / drug effects
  • Dose-Response Relationship, Drug
  • Humans
  • Kinetics
  • Leukemia, Promyelocytic, Acute
  • Microtubules / drug effects
  • Mitosis / drug effects
  • Nucleosomes / drug effects
  • Nucleosomes / pathology
  • Paclitaxel / analogs & derivatives*
  • Paclitaxel / metabolism
  • Paclitaxel / toxicity*
  • Taxoids*
  • Tumor Cells, Cultured
  • Tumor Stem Cell Assay

Substances

  • DNA, Neoplasm
  • Nucleosomes
  • Taxoids
  • 6-hydroxytaxol
  • Paclitaxel