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胃癌中的DNA损伤与修复——与hOGG1和RAD51基因多态性的相关性

DNA damage and repair in gastric cancer--a correlation with the hOGG1 and RAD51 genes polymorphisms.

作者信息

Poplawski Tomasz, Arabski Michal, Kozirowska Dorota, Blasinska-Morawiec Maria, Morawiec Zbigniew, Morawiec-Bajda Alina, Klupińska Grazyna, Jeziorski Arkadiusz, Chojnacki Jan, Blasiak Janusz

机构信息

Department of Molecular Genetics, University of Lodz, Lodz, Poland.

出版信息

Mutat Res. 2006 Oct 10;601(1-2):83-91. doi: 10.1016/j.mrfmmm.2006.06.002. Epub 2006 Jul 14.

Abstract

The cell's susceptibility to mutagens and its ability to repair DNA lesions are important for cancer induction, promotion and progression. Both the mutagens' sensitivity and the efficacy of DNA repair may be affected by variation in several genes, including DNA repair genes. The hOGG1 gene encodes glycosylase of base excision repair and RAD51 specifies a key protein in homologues recombination repair. Both can be involved in the repair of oxidative DNA lesions, which can contribute to stomach cancer. In the present work we determined the level of basal and oxidative DNA damage and the kinetics of removal of DNA damage induced by hydrogen peroxide in peripheral blood lymphocytes of 30 gastric cancer patients and 30 healthy individuals. The metrics from DNA damage and repair study were correlated with the genotypes of common polymorphisms of the hOGG1 and RAD51 genes: a G-->C transversion at 1245 position of the hOGG1 gene producing a Ser-->Cys substitution at the codon 326 (the Ser326Cys polymorphism) and a G-->C substitution at position 135 (5'-untranslated region) of the RAD51 gene (the G135C polymorphism). DNA damage and repair were evaluated by alkaline single cell gel electrophoresis (comet assay) assisted by DNA repair enzymes: endonuclease III (Nth) and formamidopyrimidine-DNA glycosylase (Fpg), preferentially recognizing oxidized DNA bases. The genotypes of the polymorphism were determined by restriction fragment length polymorphism PCR. We observed a strong association between gastric cancer occurrence, impaired DNA repair in human lymphocytes and the G/C genotype of the G135C polymorphism of the RAD51 gene. Moreover, there was a strong correlation between that genotype and stomach cancer occurrence in subjects with high level of oxidatively damaged DNA. We did not observe any correlation between the Ser1245Cys polymorphism of the hOGG1 gene and gastric cancer, including subjects with impaired DNA repair and/or high levels of endogenous oxidative DNA lesions. Therefore, our result suggest that the G135C polymorphism of the RAD51 gene may be linked with gastric cancer by the modulation of the cellular response to oxidative stress and this polymorphism may be a useful additional marker in this disease along with the genetic or/and environmental indicators of oxidative stress.

摘要

细胞对诱变剂的敏感性及其修复DNA损伤的能力对癌症的诱发、促进和进展至关重要。诱变剂的敏感性和DNA修复的功效都可能受到包括DNA修复基因在内的多个基因变异的影响。hOGG1基因编码碱基切除修复的糖基化酶,而RAD51则是同源重组修复中的关键蛋白。二者均可参与氧化性DNA损伤的修复,而这种损伤可能导致胃癌。在本研究中,我们测定了30例胃癌患者和30例健康个体外周血淋巴细胞中基础DNA损伤和氧化性DNA损伤的水平,以及过氧化氢诱导的DNA损伤的修复动力学。DNA损伤和修复研究的指标与hOGG1和RAD51基因常见多态性的基因型相关:hOGG1基因第1245位的G→C颠换导致密码子326处的丝氨酸→半胱氨酸替代(Ser326Cys多态性),以及RAD51基因第135位(5'非翻译区)的G→C替代(G135C多态性)。通过碱性单细胞凝胶电泳(彗星试验)并借助DNA修复酶:内切酶III(Nth)和甲酰胺嘧啶-DNA糖基化酶(Fpg)来评估DNA损伤和修复,这些酶优先识别氧化的DNA碱基。通过限制性片段长度多态性PCR确定多态性的基因型。我们观察到胃癌的发生、人类淋巴细胞中DNA修复受损与RAD51基因G135C多态性的G/C基因型之间存在强烈关联。此外,在氧化性DNA损伤水平较高的受试者中,该基因型与胃癌的发生之间存在强烈相关性。我们未观察到hOGG1基因的Ser1245Cys多态性与胃癌之间存在任何相关性,包括DNA修复受损和/或内源性氧化性DNA损伤水平较高的受试者。因此,我们的结果表明,RAD51基因的G135C多态性可能通过调节细胞对氧化应激的反应与胃癌相关联,并且该多态性可能是该疾病中一个有用的额外标志物,与氧化应激的遗传或/和环境指标一起。

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