Summary
DNA repair capacity was investigated in 25 normal and XP fibroblast lines after UV damage was induced, using the following methods: colonyforming ability, unscheduled DNA synthesis, and alkaline elution (which can serve as a measure of repair-specific DNA incision). The majority of the XP fibroblast lines was derived from biopsies of patients who are at present under clinical observation by Dr. E.G. Jung (Dept. of Dermatology, Mannheim Medical School).
Colony-forming ability was determined at 12 different UV dose levels and expressed in terms of D 0. Unscheduled DNA synthesis was measured autoradio-graphically. Dose-response curves (grains per nucleus versus UV dose) were established and analyzed by linear regression. The characteristic value of G 0, defined as the linear increase in the mean number of grains per nucleus when the UV dose is multiplied by the factor e (i.e., 2.72), was derived from the slope of the regression lines. For quantitating DNA-incising activity of a cell line, DNA elution curves were determined at several UV dose levels. Plotting of the initial velocities of the elution curves versus the UV doses yielded a regression line, the slope of which was used to obtain the characteristic elution value, E 0.
A descriptive correlation of all three characteristic values, D 0, G 0 and E 0, showed that in all cell lines in which colony-forming ability and unscheduled DNA synthesis were diminished, a reduction of DNA-incising activity occurred. We conclude that this reduction accounts for both the decreased colony-forming ability and unscheduled DNA synthesis.
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Abbreviations
- XP:
-
xeroderma pigmentosum
- MeSO2OMe:
-
methyl methanesulfonate
- UV light:
-
ultraviolet light
- ara-C:
-
1-β-D-arabinofuranosyl cytosine
- (Ac)2ONFln:
-
N-acetoxy-2-acetylaminofluorene
- HEPES:
-
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
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This work was supported by the Deutsche Forschungsgemeinschaft, SFB 136
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Thielmann, H.W., Popanda, O. & Edler, L. XP patients from Germany: Correlation of colony-forming ability, unscheduled DNA synthesis and single-strand breaks after UV damage in xeroderma pigmentosum fibroblasts. J Cancer Res Clin Oncol 104, 263–286 (1982). https://doi.org/10.1007/BF00406246
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DOI: https://doi.org/10.1007/BF00406246