Association between the level of ERCC-1 expression and the repair of cisplatin-induced DNA damage in human ovarian cancer cells.
Association between the level of ERCC-1 expression and the repair of cisplatin-induced DNA damage in human ovarian cancer cells.
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发表时间：
2000-03
影响因子：
2
通讯作者：
Quentin Li;Jing Jie Yu;C. Mu;M. K. Yunmbam;D. Slavsky;C. Cross;F. Bostick-Bruton;Eddie Reed
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文献类型：
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作者：
Quentin Li;Jing Jie Yu;C. Mu;M. K. Yunmbam;D. Slavsky;C. Cross;F. Bostick-Bruton;Eddie Reed
核苷酸切除修复（NER）负责铂-DNA损伤的修复。ERCC-1是NER通路中的关键基因，没有功能性ERCC-1的细胞不能修复顺铂引起的DNA损伤。因此，本研究的目的是评估ERCC-1的表达和顺铂诱导的DNA加合物在体外人卵巢癌细胞中的修复之间的关系。将MCAS细胞暴露于顺铂1小时，分别通过北方和西方印迹法测定，ERCC-1 mRNA和ERCC-1蛋白水平增加约2倍。此外，细胞核连续试验表明，ERCC-1基因转录速率增加到与稳态ERCC-1 mRNA和蛋白质相同的程度，对顺铂治疗的反应。然而，如之前报道的，MCAS细胞中ERCC-1 mRNA、ERCC-1蛋白和ERCC-1转录物的水平比A2780/CP 70细胞中的水平低两倍。此外，通过原子吸收光谱法测量，MCAS细胞中顺铂-DNA加合物的修复也比A2780/CP 70细胞中的修复少近两倍，表明在这两种人卵巢肿瘤细胞系中ERCC-1表达水平与切除修复活性之间存在强关联。这些结果表明，ERCC-1可能是一个有用的标志物，以监测修复铂-DNA损伤的肿瘤细胞，并进一步强调，潜在的药理学方法，特异性抑制ERCC-1的表达可能会增加细胞对顺铂的敏感性。
Nucleotide excision repair (NER) is responsible for the repair of platinum-DNA lesions. ERCC-1 is a critical gene within the NER pathway, and cells without a functional ERCC-1 do not repair cisplatin-caused DNA damage. The present study was therefore designed to evaluate the relationship between the expression of ERCC-1 and the repair of cisplatin-induced DNA adducts in human ovarian cancer cells in vitro. One hour exposure of MCAS cells to cisplatin yielded an approximately two-fold increment in the levels of ERCC-1 mRNA and ERCC-1 protein, as determined, respectively, by Northern and Western blottings. In addition, nuclear run-on assay showed that ERCC-1 gene transcription rate was increased to about the same extent as steady-state ERCC-1 mRNA and protein, in response to cisplatin treatment. However, the levels of ERCC-1 mRNA, ERCC-1 protein, and ERCC-1 transcript in MCAS cells are two-fold lower than those in A2780/CP70 cells, as previously reported. Furthermore, the repair of cisplatin-DNA adducts in MCAS cells, as measured by atomic absorption spectrometry, is also nearly two-fold less than that in A2780/CP70 cells, indicating a strong association between the level of ERCC-1 expression and the activity of excision repair in these two human ovarian tumor cell lines. These results suggest that ERCC-1 may be a useful marker to monitor the repair of platinum-DNA damage in tumor cells, and further highlight that potential pharmacological approaches which specifically inhibit ERCC-1 expression may increase cellular sensitivity to cisplatin.