Suzuki Nobuyasu, Kamataki Akihisa, Yamaki Junko, Homma Yoshimi
Department of Biomolecular Science, Fukushima Medical University School of Medicine, Fukushima 960-1295, Japan.
Clin Chim Acta. 2008 Jan;387(1-2):55-8. doi: 10.1016/j.cca.2007.09.001. Epub 2007 Sep 8.
The importance of circulating DNA has been recognized since the detection of mutated oncogene products in cancer patients; however, there is little information about circulating DNA in normal human plasma. We characterized circulating DNA in normal human plasma to obtain basic information.
Circulating DNA was purified from plasma samples obtained from 10 healthy donors and examined. Purified DNA was cloned and their sequence determined and analyzed. The terminal structure was examined by a labeling method.
The DNA levels in normal plasma samples were quite low (3.6-5.0 ng/ml). All 556 clones analyzed were independent, and obtained from various chromosomes and various regions of the gene. The mean values of their length and GC content were 176 bp and 53.7%, respectively. Their 5' and 3' ends were rich in C and G, respectively, and they presented as 5' protruding forms of double-stranded DNA in plasma.
Circulating DNA in normal human plasma is derived from apoptotic cells but not from necrotic cells. Structural characteristics of the circulating DNA might be associated with their stability in plasma.
自癌症患者中检测到突变的癌基因产物以来,循环DNA的重要性已得到认可;然而,关于正常人血浆中循环DNA的信息却很少。我们对正常人血浆中的循环DNA进行了表征以获取基础信息。
从10名健康供体获得的血浆样本中纯化循环DNA并进行检测。对纯化的DNA进行克隆,测定并分析其序列。通过标记法检测末端结构。
正常血浆样本中的DNA水平相当低(3.6 - 5.0 ng/ml)。分析的所有556个克隆都是独立的,且来自不同的染色体和基因的不同区域。它们的长度平均值和GC含量分别为176 bp和53.7%。它们的5'端和3'端分别富含C和G,并且在血浆中呈现为双链DNA的5'突出形式。
正常人血浆中的循环DNA源自凋亡细胞而非坏死细胞。循环DNA的结构特征可能与其在血浆中的稳定性有关。