Measurement of DNA single-strand breaks by alkaline elution and fluorometric DNA quantification

2004 ◽  
Vol 326 (2) ◽  
pp. 146-152 ◽  
Author(s):  
Marina Goumenou ◽  
Kyriaki Machera
Keyword(s):  
Dna Quantification ◽  
Single Strand ◽  
Alkaline Elution ◽  
Strand Breaks ◽  
Single Strand Breaks

scholarly journals Intracellular DNA damage produced by a series of diacridines

1985 ◽  
Vol 226 (1) ◽  
pp. 175-182 ◽  
Author(s):  
I A G Roos ◽  
L P G Wakelin ◽  
S J Henry
Keyword(s):  
Dna Damage ◽  
Single Strand ◽  
Pulse Treatment ◽  
Alkaline Elution ◽  
Strand Breaks ◽  
L1210 Cells ◽  
Single Strand Breaks ◽  
Methylene Groups ◽  
Alkane Chain ◽  
Intercalating Agents

The intracellular DNA damage produced by a series of diacridines after a 2 h pulse treatment of L1210 cells in culture was investigated by using the alkaline-elution technique. Like other intercalating agents, diacridines produce single-strand breaks and protein-DNA links. There is a large increase in both types of damage as the alkane chain linking the two 9-aminoacridine residues is increased beyond five methylene groups, which is consistent with the previously observed change from monofunctional to bifunctional intercalation [Wakelin, Romanos, Chen, Glaubiger, Canellakis & Waring (1978) Biochemistry 17, 5057-5063]. For linker chains of less than six methylene groups these agents produce less DNA damage than does the parent 9-aminoacridine at the same drug concentration. Unlike the monofunctional intercalators previously investigated [Ross, Glaubiger & Kohn (1979) Biochim. Biophys. Acta 562, 41-50; Zwelling, Michaels, Erickson, Ungerleider, Nichols & Kohn (1981) Biochemistry 20, 6553-6563; Zwelling, Kerrigan & Michaels (1982) Cancer Res. 42, 2687-2691; Zwelling, Michaels, Kerrigan, Pommier & Kohn (1982) Biochem. Pharmacol. 31, 3261-3267], there is no correlation between the number of single-strand breaks and protein-DNA links produced by these diacridines.

Single-strand breaks or alkali-sensitive sites in the DNA of human myeloma plasma cells and chronic lymphocytic leukemia lymphocytes

1985 ◽  
Vol 63 (9) ◽  
pp. 977-981 ◽  
Author(s):  
L. Brox ◽  
A. Ng ◽  
E. Pollock ◽  
A. Khaliq ◽  
A. Belch
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Plasma Cells ◽  
Human Lymphocytes ◽  
Single Strand ◽  
Lymphocytic Leukemia ◽  
General Mechanism ◽  
Alkaline Elution ◽  
Strand Breaks ◽  
Single Strand Breaks

Alkaline-elution studies showed significant levels of either DNA single-strand breaks or alkali-sensitive sites in the plasma cells of six out of six myeloma patients and in the lymphocytes of two out of four patients with chronic lymphocytic leukemia as compared with normal human lymphocytes. The increased rate of DNA elution was variable from sample to sample with a range that would correspond to that observed with 100–1000 rad (1 rad = 10 mGy) of X-ray irradiation. This alteration in DNA structure was observed in both new and advanced patients, did not appear to be related to prior therapy, and did not affect the in vitro viability of these cells. Repetitive alkaline-elution profiles obtained with tumor cells from three patients were similar on subsequent samples obtained 1 month apart. Altered DNA elution was not evident in peripheral blood lymphocytes from myeloma patients with altered plasma cell DNA elution. These observations are interesting in light of the recent hypothesis that breaking and rejoining of DNA, regulated by poly(ADP-ribosyl)ation, may be a general mechanism of altering gene expression during differentiation.

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