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a Institute of Chemical Biology and Fundamental Medicine, Siberian Division of the Russian Academy of Sciences, 8, Lavrentiev Ave., Novosibirsk 630090, Russia
Key Words: circulating DNA • deoxyribonuclease activity • amplicon-based immunoassay • blood plasma • cancer
Address for correspondence: Pavel P. Laktionov, Institute of Chemical Biology and Fundamental Medicine, Siberian Division of the Russian Academy of Sciences, 8, Lavrentiev Ave., Novosibirsk 630090, Russia. Voice: +7-383-3304654; fax: +7-383-3333677. e-mail: lakt{at}niboch.nsc.ru
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.
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