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Circulating DNA and DNase Activity in Human Blood
Author(s) -
TAMKOVICH SVETLA.,
CHEREPANOVA ANNA V.,
KOLESNIKOVA ELENA V.,
RYKOVA ELENA Y.,
PYSHNYI DMITRII V.,
VLASSOV VALENTIN V.,
LAKTIONOV PAVEL P.
Publication year - 2006
Publication title -
annals of the new york academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.712
H-Index - 248
eISSN - 1749-6632
pISSN - 0077-8923
DOI - 10.1196/annals.1368.026
Subject(s) - deoxyribonuclease i , chemistry , microbiology and biotechnology , dna , amplicon , biotinylation , deoxyribonuclease , stomach , blood plasma , biochemistry , chromatography , polymerase chain reaction , biology , gene , base sequence
The concentration of circulating DNA (cirDNA) and deoxyribonuclease activity in blood plasma of healthy donors and patients with colon or stomach cancer were analyzed. The concentration of DNA was measured using Hoechst 33258 fluorescent assay after the isolation by the glass–milk protocol. A 1‐kbp PCR product labeled with biotinylated forward and fluorescein‐labeled reverse primers was used as a substrate for DNase. DNase activity was estimated from the data of immunochemical detection of the nonhydrolyzed amplicon. The average concentration of cirDNA in the plasma of healthy donors was low (34 ± 34 ng/mL), and was accompanied with high DNase activity (0.356 ± 0.410 U/mL). The increased concentrations of cirDNA in blood plasma of patients with colon and stomach cancer were accompanied by a decrease in DNase activity below the detection level of the assay. The data obtained demonstrate that low DNase activity in blood plasma of cancer patients can cause an increase in the concentration of cirDNA.