Detection of centromeres in vinblastine- and radiation-induced micronuclei of human lymphocytes using FISH with an alpha satellite pancentromeric DNA probe.
Fluorescence in situ hybridisation (FISH) with a human alphoid satellite pancentromeric DNA probe was used to detect centromeres in micronuclei of human lymphocytes induced by gamma irradiation and by Vinblastine sulfate. In a cytokinesis-block micro-nucleus assay a dose-dependent increase of micronuclei was detected for both agents. 72-89% of vinblastine-induced micronuclei, but only 7-48% of radiation-induced micronuclei showed centromere-positive fluorescence signals. Vinblastine treatment frequencies of centromere-negative micronuclei did not increase compared to control values, nor did frequencies of centromere-positive micronuclei in irradiated lymphocytes. Since FISH with an alpha satellite DNA probe allows the direct detection of centromeric DNA sequences the spindle damaging or clastogenic effectiveness of a compound can be easily and reliably examined in a cytokinesis-block micronucleus assay in human lymphocytes.
Huber
R
R
Institute für Strahlenbiologie, GSF-Forschungszentrum für Umwelt und Gesundheit, Oberschleissheim, Germany.
Salassidis
K
K
Kulka
U
U
Braselmann
H
H
Bauchinger
M
M
D000328:Adult; D000782:Aneuploidy; D000972:Antineoplastic Agents, Phytogenic / Q000633:toxicity*; D002503:Centromere / Q000187:drug effects / Q000528:radiation effects; D015342:DNA Probes; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D016650:Fluorescein-5-isothiocyanate; D005720:Gamma Rays / Q000009:adverse effects; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008214:Lymphocytes / Q000187:drug effects / Q000528:radiation effects / Q000648:ultrastructure*; D008297:Male; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects / Q000528:radiation effects / Q000648:ultrastructure*; D008856:Microscopy, Fluorescence; D014747:Vinblastine / Q000633:toxicity*
D000972:Antineoplastic Agents, Phytogenic; D015342:DNA Probes; D014747:Vinblastine; D016650:Fluorescein-5-isothiocyanate