Varanasi U, Reichert W L, Stein J E
Environmental Conservation Division, Northwest and Alaska Fisheries Center, National Oceanic and Atmospheric Administration, Seattle, Washington 98112.
Cancer Res. 1989 Mar 1;49(5):1171-7.
The 1-butanol adduct enhancement version of the 32P-postlabeling assay was used to measure the levels of hepatic DNA adducts in the marine flatfish, English sole (Parophrys vetulus), sampled from the Duwamish Waterway and Eagle Harbor, Puget Sound, WA, where they are exposed to high concentrations of sediment-associated chemical contaminants and exhibit an elevated prevalence of hepatic neoplasms. Hepatic DNA was also analyzed from English sole from a reference area (Useless Bay, WA) and from reference English sole treated with organic-solvent extracts of sediments from the two contaminated sites. Autoradiograms of thin-layer chromatograms of 32P-labeled hepatic DNA digests from English sole from the contaminated sites exhibited up to three diagonal radioactive zones, which were not present in autoradiograms of thin-layer chromatogram maps of 32P-labeled DNA digests from English sole from the reference site. These diagonal radioactive zones contained several distinct spots as well as what appeared to be multiple overlapping adduct spots. The levels (nmol of adducts/mol of nucleotides) of total DNA adducts for English sole from Duwamish Waterway and Eagle Harbor were 26 +/- 28 (DS) and 17 +/- 9.6, respectively. All autoradiograms of DNA from fish from the contaminated sites exhibited a diagonal radioactive zone where DNA adducts of chrysene, benzo(a)pyrene, and dibenz(a,h)anthracene, formed in vitro using English sole hepatic microsomes, were shown to chromatograph. English sole treated with extracts of the contaminated sediments had adduct profiles generally similar to those for English sole from the respective contaminated sites. The chromatographic characteristics of the adducts and the similarities in adduct profiles between field-sampled English sole and those treated with contaminated sediment extracts suggested that hydrophobic aromatic compounds of anthropogenic origin were adducted to hepatic DNA of sole from contaminated sites, but not in sole from the reference site. Moreover, an initial study with winter flounder (Pseudopleuronectes americanus) from Boston Harbor, MA, an area where high concentrations of sediment-associated chemicals are present, revealed a pattern of hepatic DNA-adducts (9.0 +/- 7.8 nmol of adducts/mol of nucleotides) similar to that observed for English sole from Eagle Harbor.
采用32P后标记法的1-丁醇加合物增强版，来测量从华盛顿州普吉特海湾的杜瓦米什水道和鹰港采集的海产比目鱼——英国鲽（Parophrys vetulus）肝脏中DNA加合物的水平。在这些区域，英国鲽暴露于高浓度的与沉积物相关的化学污染物中，且肝脏肿瘤患病率升高。还分析了来自参考区域（华盛顿州无用湾）的英国鲽以及用来自两个污染场地的沉积物的有机溶剂提取物处理过的对照英国鲽的肝脏DNA。来自污染场地的英国鲽的32P标记肝脏DNA消化产物的薄层色谱图的放射自显影片显示出多达三个对角线放射性区域，而来自参考场地的英国鲽的32P标记DNA消化产物的薄层色谱图的放射自显影片中则不存在这些区域。这些对角线放射性区域包含几个不同的斑点以及似乎是多个重叠的加合物斑点。来自杜瓦米什水道和鹰港的英国鲽的总DNA加合物水平（每摩尔核苷酸的加合物纳摩尔数）分别为26±28（DS）和17±9.6。来自污染场地的鱼的所有DNA放射自显影片都显示出一个对角线放射性区域，在该区域，使用英国鲽肝脏微粒体在体外形成的芘、苯并(a)芘和二苯并(a,h)蒽的DNA加合物被显示出进行了色谱分离。用污染沉积物提取物处理的英国鲽的加合物图谱通常与来自各自污染场地的英国鲽的加合物图谱相似。加合物的色谱特征以及现场采样的英国鲽与用污染沉积物提取物处理的英国鲽之间加合物图谱的相似性表明，人为来源的疏水性芳香族化合物与来自污染场地的鲽的肝脏DNA发生了加合，但参考场地的鲽则没有。此外，对来自马萨诸塞州波士顿港的冬季比目鱼（Pseudopleuronectes americanus）的初步研究表明，该区域存在高浓度的与沉积物相关的化学物质，其肝脏DNA加合物模式（每摩尔核苷酸9.0±7.8纳摩尔加合物）与在鹰港的英国鲽中观察到的模式相似。
