Köberle Beate, Roginskaya Vera, Zima Karen S, Masters John R W, Wood Richard D
Department of Pharmacology, University of Pittsburgh Medical School and University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania, USA.
Mol Carcinog. 2008 Aug;47(8):580-6. doi: 10.1002/mc.20418.
Most testicular germ cell tumors are curable using cisplatin-based chemotherapy, and cell lines from these tumors are unusually sensitive to cisplatin and other DNA-damaging agents. It has been suggested that this might be caused by a lower-than normal nucleotide excision repair (NER) activity. Previous studies found that cell lines from testicular germ cell tumors have on average about one-third the level of the NER protein XPA in comparison to cell lines from other tumors. We asked whether over-expression of XPA protein would alleviate the cellular sensitivity and increase the DNA repair capacity of a testis tumor cell line. Increasing XPA levels in 833K cells by 10-fold did not increase resistance to UV irradiation. XPA was localized to the cell nucleus in all cell lines, before and after exposure to UV-radiation. 833K cells were proficient in removing UV radiation-induced photoproducts from the genome and increased XPA did not enhance the rate of removal. Further, over-expressing functional XPA protein did not correlate with increased resistance of 833K testis tumor cells to cisplatin. Thus, although the amount of XPA in this testis tumor cell line is lower than normal, it is sufficient for NER in vivo. The relative sensitivity of testis tumor cells to cisplatin, UV radiation, and other DNA damaging agents is likely related not to NER capacity, but to other factors such as the integrity of the p53 pathway in these cells.
大多数睾丸生殖细胞肿瘤使用基于顺铂的化疗方法可治愈,并且来自这些肿瘤的细胞系对顺铂和其他DNA损伤剂异常敏感。有人提出,这可能是由于核苷酸切除修复(NER)活性低于正常水平所致。先前的研究发现,与来自其他肿瘤的细胞系相比,来自睾丸生殖细胞肿瘤的细胞系中NER蛋白XPA的水平平均约为其三分之一。我们询问XPA蛋白的过表达是否会减轻细胞敏感性并提高睾丸肿瘤细胞系的DNA修复能力。将833K细胞中的XPA水平提高10倍并不会增加对紫外线照射的抗性。在暴露于紫外线辐射之前和之后,XPA在所有细胞系中均定位于细胞核。833K细胞能够有效地从基因组中去除紫外线辐射诱导的光产物,而增加的XPA并没有提高去除率。此外,过表达功能性XPA蛋白与833K睾丸肿瘤细胞对顺铂的抗性增加无关。因此,尽管该睾丸肿瘤细胞系中XPA的量低于正常水平,但它在体内足以进行NER。睾丸肿瘤细胞对顺铂、紫外线辐射和其他DNA损伤剂的相对敏感性可能与NER能力无关,而是与这些细胞中p53途径的完整性等其他因素有关。