Effects of antileukemia agents on nuclear matrix-bound DNA replication in CCRF-CEM leukemia cells

The effects of various antileukemic agents on DNA replication associated with the nuclear matrix were investigated in CCRF-CEM leukemia cells. Residual nuclear matrices were prepared by sequential treatment of nuclei with 1.5 M NaCl, DNase I, and Triton X-100 and contained 1-5, 10, and 37% of the to...

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Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 1988-04, Vol.48 (7), p.1850-1855
Main Authors: FERNANDES, D. J, SMITH-NANNI, C, PAFF, M. T, NEFF, T.-A. M
Format: Article
Language:English
Subjects:
Amsacrine - pharmacology
Antineoplastic agents
Antineoplastic Agents - pharmacology
Biological and medical sciences
Cell Nucleus - physiology
Cell Nucleus - ultrastructure
Chromatin - metabolism
Cytarabine - pharmacology
DNA Replication - drug effects
General aspects
Hydroxyurea - pharmacology
Medical sciences
Microscopy, Electron
Nuclear Proteins - analysis
Pharmacology. Drug treatments
Teniposide - pharmacology
Tumor Cells, Cultured
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Summary:The effects of various antileukemic agents on DNA replication associated with the nuclear matrix were investigated in CCRF-CEM leukemia cells. Residual nuclear matrices were prepared by sequential treatment of nuclei with 1.5 M NaCl, DNase I, and Triton X-100 and contained 1-5, 10, and 37% of the total nuclear DNA, protein, and phospholipid, respectively. In control cells pulse-labeled for 45 s with [3H]thymidine, the specific activity of nascent DNA was four-fold greater in the nuclear matrix fraction relative to the specific activity of the high salt-soluble (nonmatrix) DNA fraction. Pulse-labeling and reconstitution experiments indicated that this enrichment of newly replicated DNA on the nuclear matrix did not result from aggregation of nascent DNA with the matrix. A 2-h incubation of tumor cells with either 0.1 microM teniposide (VM-26), 0.2 microM VM-26, or 0.5 microM amsacrine (m-AMSA) reduced the relative specific activity of nascent DNA on the nuclear matrix by 59, 61, and 54%, respectively, compared to control cells. In contrast hydroxyurea and cytosine arabinoside, at concentrations that markedly inhibited total nuclear DNA synthesis, did not decrease the relative specific activity of newly replicated DNA on the matrix. The results provide evidence that the antiproliferative effects of the DNA topoisomerase II inhibitors, VM-26 and m-AMSA, are localized on the nuclear matrix of CCRF-CEM leukemia cells.
ISSN:0008-5472
1538-7445