Demonstration of the cell cycle positions of taxol-induced 'asters' and 'bundles' by sequential measurements of tubulin immunofluorescence, DNA content, and autoradiogaphic labeling of taxol-sensitive and -resistant cells

J. R. Roberts, E. K. Rowinsky, Ross C Donehower, J. Robertson, D. C. Allison

Research output: Contribution to journalArticle

Abstract

We used reliable and relatively inexpensive equipment to make sequential sets of measurements of antitubulin immunofluorescence, Feulgen staining, and autoradiography on the same cells. This was done to evaluate tubulin conformations, DNA content, and [3H]-thymidine incorporation in cell lines sensitive (HL60) and resistant (K562) to the novel anti-tubulin chemotherapeutic agent taxol. Numbers of cells with microtubule bundles have been found to correlate with sensitivity to taxol by clonogenic assay for several leukemic cell lines. We have found that cells with 'asters' produced by taxol exposure are in mitosis and that cells with taxol-induced 'bundles' are in G0/G1, S, and G2 phases. We further found that S-phase cells with microtubule bundles in both sensitive (HL60) and resistant (K562) cell lines were able to incorporate [3H]-thymidine after 4-hr exposure to taxol. As microtubule bundles and asters occur in cells of the same cell cycle phases in both lines, we conclude that the greater frequency of cells with microtubule bundles reported for sensitive cells after taxol treatment cannot result from drug exclusion nor from different effects of the drug on cell microtubules in these two leukemic lines.

Original languageEnglish (US)
Pages (from-to)1659-1665
Number of pages7
JournalJournal of Histochemistry and Cytochemistry
Volume37
Issue number11
StatePublished - 1989

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Tubulin
Paclitaxel
Fluorescent Antibody Technique
Cell Cycle
Microtubules
DNA
S Phase
Cell Line
Thymidine
Nucleic Acid Conformation
Cell Cycle Resting Phase
K562 Cells
G2 Phase
G1 Phase
Autoradiography
Mitosis
Pharmaceutical Preparations
Cell Count
Staining and Labeling
Equipment and Supplies

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

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title = "Demonstration of the cell cycle positions of taxol-induced 'asters' and 'bundles' by sequential measurements of tubulin immunofluorescence, DNA content, and autoradiogaphic labeling of taxol-sensitive and -resistant cells",
abstract = "We used reliable and relatively inexpensive equipment to make sequential sets of measurements of antitubulin immunofluorescence, Feulgen staining, and autoradiography on the same cells. This was done to evaluate tubulin conformations, DNA content, and [3H]-thymidine incorporation in cell lines sensitive (HL60) and resistant (K562) to the novel anti-tubulin chemotherapeutic agent taxol. Numbers of cells with microtubule bundles have been found to correlate with sensitivity to taxol by clonogenic assay for several leukemic cell lines. We have found that cells with 'asters' produced by taxol exposure are in mitosis and that cells with taxol-induced 'bundles' are in G0/G1, S, and G2 phases. We further found that S-phase cells with microtubule bundles in both sensitive (HL60) and resistant (K562) cell lines were able to incorporate [3H]-thymidine after 4-hr exposure to taxol. As microtubule bundles and asters occur in cells of the same cell cycle phases in both lines, we conclude that the greater frequency of cells with microtubule bundles reported for sensitive cells after taxol treatment cannot result from drug exclusion nor from different effects of the drug on cell microtubules in these two leukemic lines.",
author = "Roberts, {J. R.} and Rowinsky, {E. K.} and Donehower, {Ross C} and J. Robertson and Allison, {D. C.}",
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AU - Roberts, J. R.

AU - Rowinsky, E. K.

AU - Donehower, Ross C

AU - Robertson, J.

AU - Allison, D. C.

PY - 1989

Y1 - 1989

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AB - We used reliable and relatively inexpensive equipment to make sequential sets of measurements of antitubulin immunofluorescence, Feulgen staining, and autoradiography on the same cells. This was done to evaluate tubulin conformations, DNA content, and [3H]-thymidine incorporation in cell lines sensitive (HL60) and resistant (K562) to the novel anti-tubulin chemotherapeutic agent taxol. Numbers of cells with microtubule bundles have been found to correlate with sensitivity to taxol by clonogenic assay for several leukemic cell lines. We have found that cells with 'asters' produced by taxol exposure are in mitosis and that cells with taxol-induced 'bundles' are in G0/G1, S, and G2 phases. We further found that S-phase cells with microtubule bundles in both sensitive (HL60) and resistant (K562) cell lines were able to incorporate [3H]-thymidine after 4-hr exposure to taxol. As microtubule bundles and asters occur in cells of the same cell cycle phases in both lines, we conclude that the greater frequency of cells with microtubule bundles reported for sensitive cells after taxol treatment cannot result from drug exclusion nor from different effects of the drug on cell microtubules in these two leukemic lines.

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