IMPROVED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF 8-METHOXYPSORALEN MONOADDUCTS AND CROSS-LINKS IN POLYNUCLEOTIDE, DNA, AND CELLULAR-SYSTEMS - ANALYSIS OF SPLIT-DOSE PROTOCOLS
IMPROVED HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ANALYSIS OF 8-METHOXYPSORALEN MONOADDUCTS AND CROSS-LINKS IN POLYNUCLEOTIDE, DNA, AND CELLULAR-SYSTEMS - ANALYSIS OF SPLIT-DOSE PROTOCOLS
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DOI：
10.1111/j.1751-1097.1993.tb02953.x
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发表时间：
1993-06-01
影响因子：
3.3
通讯作者：
GASPARRO, FP
中科院分区：
文献类型：
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作者：
OLACK, G;GATTOLIN, P;GASPARRO, FP
用高效液相色谱和闪烁分析法测定了在不同辐照条件下经[H-3]8-甲氧基芘处理的多核苷酸、小牛胸腺DNA和哺乳动物细胞中8-甲氧基芘-胸苷光加合物的分布。分次剂量方案，与8-甲氧基补骨脂素和低剂量的长波长UV辐射处理的样品，以产生单加合物，洗涤，以除去未反应的8-甲氧基补骨脂素，然后进一步照射，以转换的单加合物的交联，进行了检查。在第一步中的光加合物的分布是依赖于UVA剂量和辐射的波长，但它是相对独立的8-甲氧基芘浓度。低能量密度和较长波长主要产生4 '，5'-单加合物，而较高能量密度和较短波长产生更多的交联。第二照射步骤将4 '，5'-单加合物转化为交联以及3，4-单加合物。在第二照射步骤之后的交联的总产率不依赖于第一步骤中使用的波长。细胞研究表明，分次给药方案适用于哺乳动物系统。这些结果可能会影响基于分次给药方案的诱变研究的解释，因为第二步可将4 '，5'-单加合物转化为预期的交联3，4-单加合物。因此，将分次给药研究第二步后致突变性增加仅与交联形成相关的解释可能需要重新检查。
The distribution of 8-methoxypsoralen-thymidine photoadducts from polynucleotides, calf thymus DNA and mammalian cells treated with [H-3]8-methoxypsoralen under a variety of irradiation conditions was determined using high-performance liquid chromatography and scintillation analysis. The split-dose protocol, with samples treated with 8-methoxypsoralen and low doses of long-wavelength UV radiation to generate monoadducts, washed to remove unreacted 8-methoxypsoralen, then irradiated further to convert the monoadducts to cross-links, was examined. The photoadduct distribution in the first step is dependent upon the UVA dose and the wavelength of the radiation, but it is relatively independent of 8-methoxypsoralen concentration. Low fluence and longer wavelengths generate mainly 4',5'-monoadducts, whereas higher fluences and shorter wavelengths yield more cross-links. The second irradiation step converts the 4',5'-monoadducts to cross-links as well as to 3,4-monoadducts. The overall yield of cross-links after the second irradiation step is not dependent upon the wavelength used in the first step. Cellular studies demonstrated that the split-dose protocol is applicable to mammalian systems. These results may affect the interpretation of mutagenesis studies based on the split-dose protocol, because the second step can convert 4',5'-monoadducts to both 3,4-monoadducts, the expected cross-links. Therefore, interpretations that link increases in mutagenicity after the second step in a split-dose study solely to cross-link formation may need re-examination.