Circulating DNA and DNase activity in human blood
Circulating DNA and DNase activity in human blood
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DOI：
10.1196/annals.1368.026
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发表时间：
2006-01-01
期刊：
影响因子：
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通讯作者：
Laktionov, Pavel P.
中科院分区：
文献类型：
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作者：
Tamkovich, Svetlana N.;Cherepanova, Anna V.;Laktionov, Pavel P.
分析了健康献血员和结肠癌、胃癌患者血浆中循环DNA（cirDNA）浓度和脱氧核糖核酸酶活性。在通过玻璃-牛奶方案分离后，使用Hoechst 33258荧光测定法测量DNA的浓度。使用生物素标记的正向引物和荧光素标记的反向引物标记的1 kbp PCR产物作为DNA酶的底物。从非水解扩增子的免疫化学检测数据估计DNA酶活性。健康供体血浆中cirDNA的平均浓度较低（34 +/- 34 ng/mL），并伴有高DNA酶活性（0.356 +/- 0.410 U/mL）。结肠癌和胃癌患者血浆中cirDNA浓度的增加伴随着DNA酶活性的降低，低于测定的检测水平。获得的数据表明，癌症患者血浆中的低DNA酶活性可导致cirDNA浓度的增加。
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.