Carcinogenesis, Vol 18, 825-832, Copyright © 1997 by Oxford University Press
H Fung, YW Kow, B Van Houten and BT Mossman
Oxidative damage is a proposed mechanism of asbestos-induced
carcinogenesis, but the detection of oxidative DNA lesions in target cells
of asbestos-induced mesothelioma has not been examined. In studies here,
DNA was isolated from both rat pleural mesothelial (RPM) cells and a human
mesothelial cell line (MET5A) after exposure in vitro to crocidolite
asbestos at various concentrations. DNA was then examined for formation of
8-hydroxydeoxyguanosine (8-OHdG) at 24, 48 and 72 h using HPLC with
electrochemical detection. In addition, steady- state mRNA levels of
manganese-containing superoxide dismutase (MnSOD) were assessed as an
indication of oxidative stress. Whereas RPM cells showed dose-dependent and
significant increases in 8-OHdG formation in response to crocidolite
asbestos or iron-chelated crocidolite fibers (but not after exposure to
glass beads), MET5A cells showed decreases in 8-OHdG. Both cell types
exhibited elevations in message levels of MnSOD. In comparison with human
MET5A cells, RPM cells exhibited increased cytotoxicity and apoptosis in
response to asbestos, as documented by cell viability assays and flow
cytometry analysis using propidium iodide. Results in RPM cells indicate
that asbestos causes oxidative damage that may result in potentially
mutagenic lesions in DNA and/or apoptosis, despite compensatory increases
in expression of an antioxidant enzyme.
ARTICLES
Patterns of 8-hydroxydeoxyguanosine formation in DNA and indications of oxidative stress in rat and human pleural mesothelial cells after exposure to crocidolite asbestos
Department of Pathology, University of Vermont, Burlington 05405, USA.
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