Mutational spectrum and genotoxicity of the major lipid peroxidation product, trans-4-hydroxy-2-nonenal, induced DNA adducts in nucleotide excision repair-proficient and -deficient human cells
Mutational spectrum and genotoxicity of the major lipid peroxidation product, trans-4-hydroxy-2-nonenal, induced DNA adducts in nucleotide excision repair-proficient and -deficient human cells
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DOI：
10.1021/bi034431g
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发表时间：
2003-07-01
期刊：
影响因子：
2.9
通讯作者：
Tang, MS
中科院分区：
文献类型：
--
作者：
Feng, ZH;Hu, WW;Tang, MS
反式-4-羟基-2-壬烯醛（4-HNE）是脂质过氧化反应的主要产物，能与DNA作用形成6-（1-羟基己基）-8-羟基-1，N-2-丙-2 '-脱氧鸟苷（4-HNE-dG）加合物，但其遗传毒性和致突变性尚不清楚。据报道，4-HNE处理人细胞可诱导p53基因密码子249（AGG*）第三个碱基发生高频率的G-C至T-A突变，这是人类癌症，特别是肝细胞癌中的突变热点。然而，密码子249处的G-C至T-A颠换被认为是由4-HNE的内源性代谢物2，3-环氧-4-羟基壬醛诱导的乙烯基-DNA加合物引起的。我们最近发现4-HNE优先在p53基因中包括密码子249（GAGG*C）的GAGG*C/A序列处形成4-HNE-dG加合物。我们的发现支持了密码子249处的G-C至T-A突变可能由4-HNE-dG加合物诱导的可能性。为了研究这种可能性，我们确定了在人细胞中复制的穿梭载体pSP 189的supF基因中由4-HNE-dG加合物诱导的突变谱。我们已经发现4-HNE-dG加合物在人类细胞中具有致突变性和遗传毒性，并且G-C至T-A颠换是由4-HNE-dG加合物诱导的最普遍的突变。此外，4-HNE-dG加合物在核苷酸切除修复（NER）缺陷的人和大肠杆菌细胞中诱导的遗传毒性和致突变性水平显著高于在NER熟练的细胞中，表明NER是修复人和大肠杆菌中4-HNE-dG加合物的主要途径。大肠杆菌细胞。总之，这些结果表明，4-HNE-dG加合物可能极大地促进了p53基因密码子249处的G-C至T-A突变，并且可能在癌发生中起重要作用。
trans-4-Hydroxy-2-nonenal (4-HNE), a major product of lipid peroxidation, is able to interact with DNA to form 6-(1-hydroxyhexanyl)-8-hydroxy-1,N-2-propano-2'-deoxyguanosine (4-HNE-dG) adducts, but its genotoxicity and mutagenicity remain elusive. It has been reported that 4-HNE treatment in human cells induces a high frequency of G-C to T-A mutations at the third base of codon 249 (AGG*) of the p53 gene, a mutational hot spot in human cancers, particularly in hepatocellular carcinoma. This G-C to T-A transversion at codon 249, however, has been thought to be caused by etheno-DNA adducts induced by the endogenous metabolite of 4-HNE, 2,3-epoxy-4-hydroxynonanal. We have recently found that 4-HNE preferentially forms 4-HNE-dG adducts at the GAGG*C/A sequence in the p53 gene including codon 249 (GAGG*C). Our finding supports the possibility that G-C to T-A mutations at codon 249 may be induced by 4-HNE-dG adducts. To investigate this possibility, we determined the mutational spectrum induced by 4-HNE-dG adducts in the supF gene of shuttle vector pSP189 replicated in human cells. We have found that 4-HNE-dG adducts are mutagenic and genotoxic in human cells, and that G-C to T-A transversions are the most prevalent mutations induced by 4-HNE-dG adducts. Furthermore, 4-HNE-dG adducts induce a significantly higher level of genotoxicity and mutagenicity in nucleotide excision repair (NER)-deficient human and Escherichia coli cells than in NER-proficient cells, indicating that NER is a major pathway for repairing 4-HNE-dG adducts in both human and E. coli cells. Together, these results suggest that 4-HNE-dG adducts may contribute greatly to the G-C to T-A mutation at codon 249 of the p53 gene, and may play an important role in carcinogenesis.