Lomonaco Stephanie L, Xu Xiaoxin S, Wang Gan
Institute of Environmental Health Sciences (IEHS), Wayne State University, Detroit, Michigan 48201, USA.
DNA Cell Biol. 2009 Jun;28(6):285-94. doi: 10.1089/dna.2008.0815.
Many anticancer drugs target the genomic DNA of cancer cells by generating DNA damage and inducing apoptosis. DNA repair protects cells against DNA damage-induced apoptosis. Although the mechanisms of DNA repair and apoptosis have been extensively studied, the mechanism by which DNA repair prevents DNA damage-induced apoptosis is not fully understood. We studied the role of the antiapoptotic Bcl-x(L) protein in nucleotide excision repair (NER)-facilitated cell protection against cisplatin-induced apoptosis. Using both normal human fibroblasts (NF) and NER-defective xeroderma pigmentosum group A (XPA) and group G (XPG) fibroblasts, we demonstrated that a functional NER is required for cisplatin-induced transcription of the bcl-x(l) gene. The results obtained from our Western blots revealed that the cisplatin treatment led to an increase in the level of Bcl-x(L) protein in NF cells, but a decrease in the level of Bcl-x(L) protein in both XPA and XPG cells. The results of our immunofluorescence staining indicated that a functional NER pathway was required for cisplatin-induced translocation of NF-kappaB p65 from cytoplasm into nucleus, indicative of NF-kappaB activation. Given the important function of NF-kappaB in regulating transcription of the bcl-x(l) gene and the Bcl-x(L) protein in preventing apoptosis, these results suggest that NER may protect cells against cisplatin-induced apoptosis by activating NF-kappaB, which further induces transcription of the bcl-x(l) gene, resulting in an accumulation of Bcl-x(L) protein and activation of the cell survival pathway that leads to increased cell survival under cisplatin treatment.
许多抗癌药物通过产生DNA损伤并诱导细胞凋亡来靶向癌细胞的基因组DNA。DNA修复可保护细胞免受DNA损伤诱导的凋亡。尽管DNA修复和凋亡的机制已得到广泛研究,但DNA修复阻止DNA损伤诱导凋亡的机制尚未完全清楚。我们研究了抗凋亡Bcl-x(L)蛋白在核苷酸切除修复(NER)促进细胞抵抗顺铂诱导凋亡中的作用。使用正常人成纤维细胞(NF)以及NER缺陷的A型着色性干皮病(XPA)和G型着色性干皮病(XPG)成纤维细胞,我们证明顺铂诱导的bcl-x(l)基因转录需要功能性NER。我们的蛋白质免疫印迹结果显示,顺铂处理导致NF细胞中Bcl-x(L)蛋白水平升高,但在XPA和XPG细胞中Bcl-x(L)蛋白水平降低。我们的免疫荧光染色结果表明,顺铂诱导的NF-κB p65从细胞质转运到细胞核需要功能性NER途径,这表明NF-κB被激活。鉴于NF-κB在调节bcl-x(l)基因转录中的重要功能以及Bcl-x(L)蛋白在防止凋亡中的作用,这些结果表明NER可能通过激活NF-κB来保护细胞免受顺铂诱导的凋亡,NF-κB进而诱导bcl-x(l)基因转录,导致Bcl-x(L)蛋白积累并激活细胞存活途径,从而使细胞在顺铂处理下存活率增加。