ERCC2/XPD Lys751G1n alter DNA repair efficiency of platinum-induced DNA damage through P53 pathway

作者：Zhang, Guopei; Guan, Yangyang; Zhao, Yuejiao; van der Straaten, Tahar; Xiao, Sha; Xue, Ping; Zhu, Guolian; Liu, Qiufang; Cai, Yuan; Jin, Cuihong; Yang, Jinghua; Wu, Shengwen; Lu, Xiaobo^*

来源：Chemico-Biological Interactions, 2017, 263: 55-65.

DOI：10.1016/j.cbi.2016.12.015

• ERCC2/XPDLys751GInrs13181
• Nucleotide excision repair (NER)
• DNA repair capacity (DRC)
• Cisplatin resistance
• Transfected cell model

Platinum-based treatment causes Pt-DNA adducts which lead to cell death. The platinum-induced DNA damage is recognized and repaired by the nucleotide excision repair (NER) system of which ERCC2/XPD is a critical enzyme. Single nucleotide polymorphisms in ERCC2/XPD have been found to be associated with platinum resistance. The aim of the present study was to investigate whether ERCC2/XPD Lys751Gln (rs13181) polymorphism is causally related to DNA repair capacity of platinum-induced DNA damage. First, cDNA clones expressing different genotypes of the polymorphism was transfected to an ERCC2/XPD defective CHO cell line (UV5). Second, all cells were treated with cisplatin. Cellular survival rate were investigated by MIT growth inhibition assay, DNA damage levels were investigated by comet assay and RAD51 staining. The distribution of cell cycle and the change of apoptosis rates were detected by a flow cytometric method (FCM). Finally, P53mRNA and phospho-P53 protein levels were further investigated in order to explore a possible explanation. As expected, there was a significantly increased in viability of UV5(ERCC2) ((AA)) as compared to UV5(ERCC2(CC)) after cisplatin treatment. The DNA damage level of UV5(ERCC2) ((AA)) was significant decreased compared to UV5(ERCC2 (AA)) at 24 h of treatment. Mutation of ERCC2rs13181 AA to CC causes a prolonged S phase in cell cycle. UV5(ERCC2 (AA)) alleviated the apoptosis compared to UV5(ERCC2) ((CC)), meanwhile P53mRNA levels in UVERCC2 (AA) was also lower when compared UV5(ERCC2) ((CC)). It co-incides with a prolonged high expression of phospho-P53, which is relevant for cell cycle regulation, apoptosis, and the DNA damage response (DDR). We concluded that ERCC2/XPD rs13181 polymorphism is possibly related to the DNA repair capacity of platinum-induced DNA damage. This functional study provides some clues to clarify the relationship between cisplatin resistance and ERCC2/XPDrs13181 polymorphism.

• 出版日期2017-2-1
• 单位中国医科大学; 辽宁省肿瘤医院