title: ERCC2/XPD Lys751Gln alter DNA repair efficiency of platinum-induced DNA damage through P53 pathway.
abstract: 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 MTT 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 UV5ERCC2 (AA)as compared to UV5ERCC2 (CC)after cisplatin treatment. The DNA damage level of UV5ERCC2 (AA)was significant decreased compared to UV5ERCC2 (CC)at 24 h of treatment. Mutation of ERCC2rs13181 AA to CC causes a prolonged S phase in cell cycle. UV5ERCC2 (AA)alleviated the apoptosis compared to UV5ERCC2 (CC), meanwhile P53mRNA levels in UVERCC2 (AA)was also lower when compared UV5ERCC2 (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.
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Guopei
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Yangyang
authors.initials: Y
authors.lastname: Guan
authors.affiliation: Cancer Hospital of China Medical University/Liaoning Cancer Hospital & Institute, PR China.
authors.forename: Yuejiao
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Dept. Clinical Pharmacy and Toxicology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Tahar
authors.initials: T
authors.lastname: van der Straaten
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Sha
authors.initials: S
authors.lastname: Xiao
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Ping
authors.initials: P
authors.lastname: Xue
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Guolian
authors.initials: G
authors.lastname: Zhu
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Qiufang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Cai
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Cuihong
authors.initials: C
authors.lastname: Jin
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Jinghua
authors.initials: J
authors.lastname: Yang
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China.
authors.forename: Shengwen
authors.initials: S
authors.lastname: Wu
authors.affiliation: Dept. of Toxicology, School of Public Health, China Medical University, Shenyang, PR China. Electronic address: xblu@cmu.edu.cn.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Lu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D016466:CHO Cells; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000633:toxicity; D020552:Comet Assay; D006224:Cricetinae; D003412:Cricetulus; D004249:DNA Damage / Q000187:drug effects*; D004260:DNA Repair / Q000187:drug effects*; D006801:Humans; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D010984:Platinum / Q000633:toxicity*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*; D051759:Xeroderma Pigmentosum Group D Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D010984:Platinum; D051759:Xeroderma Pigmentosum Group D Protein; D002945:Cisplatin
keywords: Cisplatin resistance; DNA repair capacity (DRC); ERCC2/XPDLys751Glnrs13181; Nucleotide excision repair (NER); Transfected cell model