Increased sensitivity of damaged DNA to digestion with nuclease S1 as assessed in single cells by flow cytometry

Journal Article · Fri Jan 01 00:00:00 EST 1993 · Cytometry (Baltimore); (United States)

DOI:https://doi.org/10.1002/cyto.990140110· OSTI ID:6657826

Prosperi, E; Supino, R; Bottiroli, G ^[1]

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1. Univ. di Pavia (Italy)

DNA sensitivity to digestion with nuclease S1 was investigated in cells irradiated with gamma rays, or treated with the antitumor drug adriamycin (Adr). The nuclease-resistant DNA fraction was determined by propidium iodide staining. Treated cells were found to be more sensitive to nuclease digestion than the undamaged controls. Gamma ray-induced strand breaks were detectable at doses up to 10 Gy; an increase in the reaction temperature, from 37[degrees] to 63[degrees]C, was necessary in order to detect higher levels of damage. Nuclease S1 sensitivity in Adr-treated cells showed a single-peak, concentration-dependent relationship, in agreement with the known self-inhibitory effect exerted by high drug doses. Determination of DNA digestion could be performed in combination with other cellular parameters (e.g., protein content). Detection of drug-resistant cells in a heterogeneous population of small-cell lung carcinoma was achieved on the basis of the different sensitivity of the cells to enzymatic digestion. These results indicate that nuclease S1 may be a useful probe for studying in single cells DNA alterations induced by drugs or radiation. 31 refs., 8 figs.

• OSTI ID: 6657826
• Journal Information: Cytometry (Baltimore); (United States), Vol. 14:1; ISSN 0196-4763
• Country of Publication: United States
• Language: English

Related Subjects

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
DNA
DIGESTION
SENSITIVITY
GAMMA RADIATION
CHEMICAL RADIATION EFFECTS
STRAND BREAKS
DETECTION
DAMAGE
DOSES
DRUGS
LEVELS
ELECTROMAGNETIC RADIATION
IONIZING RADIATIONS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
RADIATION EFFECTS
RADIATIONS
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture