© 1990 by Oxford University Press
Journal of the National Cancer Institute, Vol. 82, No. 9, 779-783,
May 2, 1990
© 1990 Oxford University Press
Detection of Etoposide Resistance by Measuring DNA Damage in Individual Chinese Hamster Cells
British Columbia Cancer Research Centre Vancouver, BC, Canada
*Correspondence to: Peggy L. Olive, Ph.D., Medical Biophysics Unit, British Columbia Cancer Research Centre, 601 W. 10th Ave., Vancouver, BC, V5Z IL3 Canada.
The comet assay, which measures DNA strand breakage in individual cells, was used to examine the relation between DNA damage, cell survival, and resistance to the topoisomerase II inhibitor etoposide (VP-16). Chinese hamster V79–171b) cells and a VP-16-resistant subline (VP) were exposed to VP-16 as monolayers or spheroids. The comet assay was comparable in sensitivity to the DNA precipitation and alkali unwinding assays for detecting DNA strand breaks induced by VP-16. However, unlike conventional DNA damage assays, the comet assay also indicated heterogeneity in cell response. For V79 multicell spheroids exposed to VP-16, the external cycling cells were 50 times more sensitive to killing and DNA damage than the internal noncycling cells; the comet assay indicated the fraction of cells resistant to the drug. VP cells, which were 10 times more resistant to killing and DNA damage by VP-16 than the parent cell line, could also be identified in mixed populations with the use of this method. These results suggest that the comet assay could be useful in predicting tumor cell response to DNA- damaging agents. [J Natl Cancer Inst 82: 779–783, 1990]
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