Capovilla Alexio, Arbuthnot Patrick
Department of Molecular Medicine and Haematology, University of the Witwatersrand Medical School, 7 York Road, 2193, Parktown, South Africa.
FEBS Lett. 2002 May 8;518(1-3):144-8. doi: 10.1016/s0014-5793(02)02686-8.
Human hepatocytes are particularly exposed to genotoxins, and nucleotide excision repair (NER) in these cells is essential for the maintenance of genome integrity. To characterize NER under conditions that closely resemble the pathway in vivo, we report the preparation and use of primary human fetal liver extracts to define the repair of a 1,3-intrastrand d(GpTpG)-cisplatin DNA lesion. Endonucleolytic cleavage at unique sites on either side of the adduct occurs at similar positions to the dominant NER incisions that have been reported for HeLa extracts. However, incisions effected by primary hepatocyte extracts are more precise as no secondary cleavage sites are detected 5' and 3' to the cisplatin lesion.
人类肝细胞特别容易受到基因毒素的影响，这些细胞中的核苷酸切除修复（NER）对于维持基因组完整性至关重要。为了在与体内途径非常相似的条件下表征NER，我们报告了原代人胎肝提取物的制备和使用，以确定1,3-链内d（GpTpG）-顺铂DNA损伤的修复情况。加合物两侧独特位点的内切核酸酶切割发生在与已报道的HeLa提取物的主要NER切口相似的位置。然而，原代肝细胞提取物产生的切口更精确，因为在顺铂损伤的5'和3'端未检测到二级切割位点。
