hOGG1基因多态性与川北地区肺癌易感性关系的研究

目的探讨四川北部地区汉族人群8-羟基鸟嘌呤糖苷酶(hOGG1)基因Ser326Cys位点多态性状况及其与该地区肺癌易感性的关系,并与其他地区人群进行比较。方法采用病例对照研究方法和聚合酶链式反应-限制性片段长度多态性(polymerise chain reaction-restriction fragment length polymorphism,PCR-RFLP)方法检测四川北部地区肺癌患者125例和非肿瘤患者125例(对照组)hOGG1基因第326位点Ser/Cys基因型分布,并比较不同基因型与肺癌发病危险的关系。结果病例组的Ser/Ser、Ser/Cys、Cys/Cys基因型频率分别为15.2%、27.2%、57.6%,而对照组分别为32.0%、42.4%、25.6%。χ2检验显示两组基因型分布差异有统计学意义(P<0.05)。与携带Ser/Ser基因型相比,携带Ser/Cys基因型者患肺癌风险增加(OR=1.351,95%CI=0.674～2.707,P=0.397),而携带Cys/Cys基因型者肺癌风险增加3倍(OR=4.737,95%CI=2.384～9.413,P=0.000)。结论 hOGG1基因Ser326Cys多态性可能与四川北部肺癌易感性有关,携带Cys/Cys基因型肺癌风险明显增加。     

8-羟基鸟嘌呤糖苷酶1基因多态性与喉癌易感性的关系

目的:探讨8-羟基鸟嘌呤糖苷酶1基因(hOGG1)Ser326Cys多态性与喉癌遗传易感性的关系.方法:采用病例-对照设计,选择72例喉鳞状细胞癌患者(病例组),并与72例性别比一致,平均年龄相差＜5岁的对照组进行比较.采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术分析hOGG1c.326多态性.结果:病例组hOGG1c.326杂合型及突变(Ser/Cys Cys/Cys)比例高于对照组(P＜0.05),携带hOGG1c.326Ser/Cys Cys/Cys基因型个体较携带hOGG1c.326Ser/Ser基因型的个体患喉癌的风险升高了2.54倍.结论:hOGG1c.326(Ser/Cys或Cys/Cys)基因型可能是喉癌发病的风险因子,其可作为一种易感性分子标志物,对喉癌的早期预防具有一定的参考价值.     

APE1和ERCC1基因多态性和肺腺癌易感性关系研究

目的探讨APE1基因和ERCC1基因多态性和肺腺癌易感性之间的关系。方法应用实时荧光定量PCR技术对200例肺腺癌患者和200例非肿瘤患者进行ERCC1C118T和APE1T141G基因多态性分析。结果肺腺癌组ERCC1 C118T TT基因型频率高于对照组,然而差异无统计学意义（P=0．35）。肺腺癌组APE1T141GGG基因型低于对照组,差异具有统计学意义（P=0．04）,与TT基因型携带者相比,GG基因型携带者患肺腺癌的发病风险显著降低（OR=0．55,95％CI=0．31—0．98）。结论APE1T141G多态性可能和肺腺癌易感性相关,GG基因型是肺腺癌的遗传保护因素。     

DNA修复基因XPD单核苷酸多态性与肺癌遗传易感性

目的探讨XPD基因单核苷酸多态性与肺癌遗传易感性的关系。方法以聚合酶链反应-限制性片段长度多态性方法分析肺癌患者(n=114)和按性别、年龄频数配比的对照者(n=114)XPD基因Asp312Asn和Lys751Gln位点的多态性,比较不同基因型与肺癌风险的关系,并探讨吸烟在其中的影响。结果与携带XPD312Asp/Asp基因型者比较,携带至少1个312Asn等位基因的个体罹患肺癌的风险增加1.55倍(95%CI1.02～2.39)。而与携带XPD751Lys/Lys基因型者比较,携带至少1个751Gln等位基因的个体罹患肺癌的风险并没有显著增加(校正OR=0.96;95%CI0.53～1.72)。交互作用分析显示,携带至少1个312Asn等位基因并吸烟者肺癌风险增加5.14倍(95%CI1.82～9.16),其中重度吸烟者肺癌风险增加高达7.32倍(95%CI2.17～21.18)。结论DNA修复基因XPD Asp312Asn多态性可能与山东地区汉族人群肺癌遗传易感性有关,并可显著增加吸烟对肺癌的危险性。     

CDC6基因G1321A多态性与肝细胞肝癌遗传易感性关联研究

目的探讨细胞分裂周期蛋白6基因（CDC6）的G1321A（rs13706）位点多态性在中国广西人群中与肝细胞肝癌（hepatecellularcarcinoma,HCC）易感性的关系。方法采用病例一对照研究方法,对580例HCC患者和667例对照者,运用聚合酶链反应一限制性片段长度多态性实验分析CDC6基因rs13706位点多态性,并应用Logistic回归模型分析该位点多态性与肝细胞肝癌遗传易感性的关系。结果携带AA基因型的个体较携带GG基因型的个体患HCC的风险显著降低[Oddratio（OR）=0．474,95％confidenceinterval（95％C／）=0．315～0．714,P=0．0009],并在分层分析中发现rs13706位点与HBV感染因素存在显著的交互作用（P_homgeneity=0．033）。结论CDC6基因rs13706位点可能与中国广西人群HCC发生相关。     

RANTES 基因多态性与结核性脓胸发病风险的关系

目的：探讨河北地区受激活调节正常 T 细胞表达和分泌因子（regulated upon activation normal T cell ex－pressed and secreted factor,RANTES）基因－403G／A 和－28C／G 单核苷酸多态性（SNP）与结核性脓胸的关系。方法采用聚合酶链反应－限制性片段长度多态性（PCR－RFLP）方法检测300例结核性脓胸患者和300例对照人群RANTES基因－403G／A 和－28C／G 的基因型。结果 RANTES 基因－403G／A SNP 基因型频率和等位基因频率在结核性脓胸组和正常组间差异有统计学意义（ P ＝0．004和 P ＝0．008）,与 GG 基因型相比,携带 AG ＋AA 基因型可增加结核性脓胸发病风险（OR ＝1．40395％,CI ＝1．015～1．939）。 RANTES 基因－28C／G SNP 基因型频率和等位基因频率在结核性脓胸组和正常组间差异无统计学意义（ P ＝0．128和 P ＝0．064）,与 CC 基因型相比,携带 CG ＋GG 基因型与结核性脓胸发病风险无关（OR ＝1．31495％,CI ＝0．891～1．936）。结论 RANTES 基因－403G／A SNP 可能与结核性脓胸的发病风险有关,即携带 AG ＋AA 基因型增加结核性脓胸发病风险;RANTES 基因－28C／G SNP 可能与结核性脓胸发病风险无关。     

CCNE1、RIP2基因多态性与膀胱癌发病风险的关系

目的 细胞周期蛋白 E1 （CCNE1） 基因和受体相互作用蛋白 2 （RIP2） 基因区域分别存在多态性位点 rs8102137 和 rs42490, 探讨这 2 种多态性与膀胱癌的发病风险及膀胱癌的病理分级和分期之间的关系。方法 收集膀胱癌患者 （膀胱癌组） 176 例及非肿瘤体检患者 （对照组） 210 例血样本并提取 DNA, 聚合酶链反应 （PCR） 测序方法检测 CCNE1 （rs8102137） 和 RIP2 （rs42490） 的多态性。根据术后病理结果确定膀胱癌患者的分级及分期。分析膀胱癌组和对照组中等位基因和基因型的分布差异。对 2 组间基因型的分布情况进行比较, 分析 CCNE1 （rs8102137） 和 RIP2 （rs42490） 各基因型与膀胱癌患者的临床资料间的关系, 及与膀胱癌遗传易感性的关系。结果 对照组样本的基因型分布具有良好的群体代表性。CCNE1 （rs8102137） 突变型 （C/T+C/C） 在膀胱癌组中频率 （40.91%） 高于对照组（30.95%, OR=1.54, 95%CI： 1.02～2.45, P＜0.05）。RIP2 （rs42490） 突变型 （A/G+G/G） 在膀胱癌组频率 （72.73%） 高于对照组 （62.38%, OR=1.61, 95%CI： 1.04～2.48, P＜0.05）。不同膀胱癌病理分级、 分期患者的 CCNE1 （rs8102137） 和 RIP2 （rs42490） 基因多态性差异无统计学意义。结论 CCNE1 （rs8102137）、 RIP2 （rs42490） 基因型与膀胱癌易感性关系密切, 携带突变型等位基因的个体患膀胱癌的风险显著高于野生型个体。     

p21和p27基因多态性与女性肺癌的关联研究

目的探讨p21和p27基因多态性与女性非小细胞肺癌（NSCLC）遗传易感性的关系。方法应用聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）方法,分析110例女性NSCLC患者和124例女性健康对照人群p21基因3＇非翻译区（3＇UTR）和p27基因第109密码子多态性位点的基因型。结果p21基因突变型（C/T＋T/T）频率在病例组（76.4%）显著高于对照组（56.5%）[（χ2=10.27,P=0.001）。携带T等位基因（C/T和T/T基因型）能显著增加这一人群NSCLC发病风险（经年龄校正的OR=2.60,95%CI（1.46,4.61）]。根据吸烟状况和年龄分层分析发现,p21基因突变型（C/T＋T/T）频率在不吸烟病例组（86.0%）和≥50岁病例组（76.6%）均显著高于对照组（P〈0.01）,携带C/T和T/T基因型可显著增高不吸烟者和50岁以上人群的NSCLC发病风险[经年龄校正的OR分别为4.95和2.68,95%CI分别为（2.36,10.40）和（1.41,5.08）]。p27等位基因和基因型总体分布在2组差异无统计学意义（P〉0.05）,分层分析也未显示出结果有差异性。对淋巴结转移的研究显示p21和p27基因多态性与NSCLC患者的淋巴结转移无关。结论p21基因3＇UTR多态性与该女性人群NSCLC发病风险有关,C/T、T/T基因型可能增加NSCLC发病风险,尤其对于不吸烟个体及50岁以上者其可能是NSCLC的发病危险因素。p27基因多态性与该女性人群NSCLC遗传易感性无关。p21和p27基因多态性与NSCLC患者淋巴结转移无关。     

IL-1F10基因多态性与类风湿关节炎遗传易感性研究

目的探讨类风湿关节炎易感性与白细胞介素-1F10（IL-1FIO）基因中rs6743376和rs6761276位点基因多态性的关系。方法采用病例对照的研究方法,收集安徽医科大学第一附属医院风湿科门诊确诊的类风湿关节炎患者184人和同期正常健康体检人群184人,采用高温连接酶的连接酶检测反应（LDR）方法检测两位点的单核苷酸多态性,分析基因型及等位基因频率在两组中的分布。结果白细胞介素-1FIO（IL-1FIO）基因中rs6743376和rs6761276位点基因型频率和等位基因频率在病例组和对照组中的分布差异无统计学意义,rs6743376位点基因型频率（x2=2．04,P=0．361）,等位基因频率（X2=0．187,P=0．667）;rs6761276位点基因型频率（X2．28,P=0．320）,等位基因频率（X2=0．206,P=0．650）。结论中国安徽省汉族类风湿关节炎患者遗传易感性与白细胞介素-1F10基因中rs6743376和rs6761276位点无关。     

ERCC2／XPD 和 XRCC1基因多态性评估食管癌预后的应用价值

目的：探讨DNA 损伤修复基因核苷酸切除修复交叉互补基因2（ERCC2／XPD ）和X线修复交叉互补基因1（XRCC1）的基因多态性评估食管癌预后的价值。方法采集100例食管癌患者（试验组）和80例正常受试者（对照组）静脉血,采用聚合酶链反应‐限制性片段长度多态性方法检测ERCC2／XPD Lys751Gln和XRCC1 Arg399Gln基因多态性,Logistic回归分析计算风险比,分析基因型与食管癌复发、转移的相关性。结果试验组和对照组变异型等位基因 ERCC2／XPD Lys751Gln频率为15.7％和13.0％;与野生型Lys／Lys相比,携带Lys／Gln和Gln／Gln基因型易患食管癌（P＜0．05）。试验组和对照组变异型等位基因XRCC1399Gln频率分别为29.8％和30.2％;与野生型Arg／Arg相比,携带Arg／Gln和Gln／Gln基因型易患食管癌（P＜0．05）。食管癌复发、转移与ERCC2／XPD基因Lys／Gln、Gln／Gln基因型及XRCC1399 Arg／Arg基因型相关（P＜0．05）,而与ERCC2／XPD Lys／Lys基因型及XRCC1399基因Arg／Gln、Gln／Gln基因型无明显相关性（ P＞0．05）。结论 ERCC2／XPD和XRCC1基因多态性可增加食管癌发生风险,影响食管癌患者的预后。     

核苷酸切除修复酶基因多态性与晚期胃癌化疗敏感性的关系研究

目的探讨核苷酸切除修复酶基因118密码子(ERCC 1118)和751密码子(XPD 751)多态性与晚期胃癌患者对5-氟尿嘧啶(5-Fu)/铂类药物化疗敏感性的关系。方法 91例患者给予5-Fu/铂类药物方案治疗,化疗前检测ERCC1 118和XPD 751基因型。观察化疗疗效与ERCC1 118和XPD 751多态性的关系。结果 本组患者化疗有效率为37.4%。ERCC1 118的3种基因型化疗有效率比较差异无统计学意义(P>0.05);XPD 751的杂合子Lys/Gln基因型的化疗有效率为11.1%,低于纯合子Lys/Lys和Gln/Gln基因型的38.0%和100.0%,差异有统计意义(P<0.01)。结论核苷酸切除修复酶基因XPD 751基因多态性与晚期胃癌对5-Fu/铂类药物化疗疗效有关。     

The ERCC2/XPD Lys751Gln polymorphism affects DNA repair of benzo[a]pyrene induced damage,tested in an in vitro model

Nucleotide excision repair (NER) is an important defense mechanism of the body to exogenous carcinogens and mutagens, such as benzo[a]pyrene (B[a]P). Genetic polymorphisms in ERCC2/XPD, a critical element in NER, are thought to be associated with individual's cancer susceptibility. Although ERCC2/XPD Lys751Gln (rs13181) is the most studied polymorphism, the impact of this polymorphism on DNA repair capacity to carcinogen remains unclear. In the present study, cDNA clones carrying different genotypes of ERCC2/XPD (Lys751Gln) were introduced into an ERCC2/XPD deficient cell line (UV5) in a well-controlled biological system. After B[a]P treatment, cell growth inhibition rates and DNA damage levels in all cells were detected respectively. As expected, we found that the DNA repair capacity in UV5 cells was restored to levels similar to wildtype parent AA8 cells upon introduction of the cDNA clone of ERCC2/XPD (Lys751). Interestingly, after B[a]P treatment, transfected cells expressing variant ERCC2/XPD (751Gln) showed an enhanced cellular sensitivity and a diminished DNA repair capacity. The wildtype genotype AA (Lys) was found to be associated with a higher DNA repair capacity as compared to its polymorphic genotype CC (Gln). These data indicate that ERCC2/XPD Lys751Gln polymorphism affects DNA repair capacity after exposure to environmental carcinogens such as B[a]P in this well-controlled in vitro system and could act as a biomarker to increase the predictive value to develop cancer.     

hOGG1 promoter methylation,hOGG1 genetic variants and their interactions for risk of coal-borne arsenicosis: A case-control study

To identify the effect of hOGG1 methylation, Ser326Cys polymorphism and their interactions on the risk of coal-borne arsenicosis, 113 coal-borne arsenicosis subjects and 55 reference subjects were recruited. Urinary arsenic contents were analyzed with ICP-MS. hOGG1 methylation and Ser326Cys polymorphism was measured by mehtylation-specific PCR and restriction fragment length polymorphism PCR in PBLCs, respectively. The results showed that the prevalence of methylated hOGG1 and variation genotype (326 ^Ser/Cys & 326 ^Cys/Cys) were increased with raised levels of urinary arsenic in arsenicosis subjects. Increased prevalence of methylated hOGG1 and variation genotype were associated with raised risk of arsenicosis. Moreover, the results revealed that variant genotype might increase the susceptibility to hOGG1 methylation. The interactions of methylated hOGG1 and variation genotype were also found to contribute to increased risk of arsenicosis. Taken together, hOGG1 hypermethylation, hOGG1 variants and their interactions might be potential biomarkers for evaluating risk of coal-borne arsenicosis.     

The human 8-oxoguanine DNA N-glycosylase 1 (hOGG1) DNA repair enzyme and its association with lung cancer risk

OBJECTIVE: The human 8-oxoguanine DNA N-glycosylase 1 gene encodes a DNA glycosylase that is involved in the base excision repair of 8-hydroxy-2-deoxyguanine from oxidatively-damaged DNA and expressed in lung tissue. The codon 326 polymorphism in the hOGG1 gene has been suggested to reduce DNA repair enzyme activity based on in vitro functional analysis. The goal of the present study is to determine whether the codon 326 polymorphism was significantly associated with alterations in individual risk for lung cancer. METHODS: To determine whether hOGG1 plays a role in risk for lung cancer, we measured the prevalence of the Ser326Cys polymorphism in incident lung cancer patients and matched non-cancer controls. hOGG1 genotyping was performed by PCR-restriction fragment length polymorphism analysis of genomic DNA isolated from 179 Caucasian lung cancer cases and 358 controls individually matched in a 1:2 ratio by race-, sex- and age (+/- 5 years). RESULTS: Significantly increased risk for lung cancer was observed for both the hOGG1 326 (odds ratio [OR] = 1.9, 95% confidence interval [CI] = 1.2-2.9) and hOGG1 326 genotypes (OR = 3.8, 95% CI = 1.4-10.6). The increased risk for lung cancer was observed for subjects with both the hOGG1 326 (OR = 1.7, 95% CI = 1.1-2.8) and hOGG1 326 genotypes (OR = 4.9, 95% CI = 1.5-16.1) in ever-smokers. A significant association was found between hOGG1 genotypes and lung cancer risk with a dose-dependent effect with smoking. Significantly increased risk for variant hOGG1 genotypes was observed for all non-small cell lung cancer patients. CONCLUSION: These results suggest that the hOGG1 Ser326Cys polymorphism plays an important role in the risk for lung cancer and is linked to exposure to tobacco smoke.     

Polymorphisms and haplotypes of DNA repair and xenobiotic metabolism genes and risk of DNA damage in Chinese vinyl chloride monomer (VCM)-exposed workers

In this case-control study, we investigated the association between DNA damage and genetic susceptibility among vinyl chloride monomer (VCM)-exposed workers. The cumulative exposure dose of VCM was calculated based on the workers' duration of exposure and the geometric mean concentration of VCM in the workplace. DNA damage to peripheral blood lymphocytes was measured by single cell gel electrophoresis (SCGE) assay, and single nucleotide-polymorphisms (SNPs) in xenobiotic metabolism and DNA repair genes were detected by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. Univariate analysis showed that the CYP2E1 c1c2/c2c2 and XPD751 Lys/Gln and Gln/Gln genotypes were significantly associated with the levels of DNA damage (P<0.01 and 0.05, respectively). Further logistic regression analysis showed a significant association between CYP2E1 c1c2/c2c2 and DNA damage, and risk of having increased levels of DNA damage was more pronounced in those individuals having XRCC1 194 mutant genotypes and/or XPD751 Lys/Gln and Gln/Gln genotypes. Although most of the XPD and XRCC1 haplotypes did not show any significant difference, the XRCC1 haplotype Trp194-Arg280 was significantly over-represented in the case group (P<0.05; OR 2.09; 95% CI: 1.07-4.06) than in controls. Overall, our data suggest that the genotypes of CYP2E1, XRCC1 194, and XPD 751 were associated with the level of DNA damage and may contribute to individual sensitivity to DNA damage induced by VCM in the workplace.     

Association of <Emphasis Type="Italic">hOGG1</Emphasis> genotype with life style and oxidative DNA damage among Chinese ethnic populations

To assess the natural variation of hOGG1 gene and the gene–environmental interactions, the hOGG1 codon 326 polymorphism and urinary 8-OHdG levels were investigated in large samples (n = 953) of healthy individuals from five Chinese ethnic populations by using PCR-RFLP and HPLC-ECD. Life-style parameters under study were obtained through a questionnaire. The allelic frequencies of the hOGG1 gene in the Chinese populations were found to be 0.16 (Ser/Ser), 0.49 (Ser/Cys) and 0.35 (Cys/Cys), respectively. The frequencies of Ser326Cys polymorphism were significantly different among the five Chinese ethnic populations (P = 0.002). No association was found between the hOGG1 gene polymorphism and other life-style parameters except for the association between Ser326Cys and smoking (P = 0.027). A significant increase of urinary 8-OHdG level was observed in Cys326Cys allelic healthy subjects (P = 0.033). These results suggest that there are natural variations of hOGG1 gene among Chinese ethnic populations. Smoking relates to Cys/Cys polymorphism frequencies, and oxidative DNA damage is repaired less in individuals with the hOGG1 Cys326Cys genotype.     

Polymorphic trial in oxidative damage of arsenic exposed Vietnamese

Arsenic causes DNA damage and changes the cellular capacity for DNA repair. Genes in the base excision repair (BER) pathway influence the generation and repair of oxidative lesions. Single nucleotide polymorphisms (SNPs) in human 8-oxoguanine DNA glycosylase (hOGG1) Ser326Cys; apurinic/apyrimidinic endonuclease (APE1) Asp148Glu; X-ray and repair and cross-complementing group 1 (XRCC1) Arg280His and Arg399Gln in the BER genes were analyzed, and the relationship between these 4 SNPs and the urinary 8-hydroxy-2′-deoxyguanosine (8-OHdG) concentrations of 100 Vietnamese population exposed to arsenic was investigated. Individuals with hOGG1 326Cys/Cys showed significantly higher urinary 8-OHdG concentrations than did those with 326 Ser/Cys and Ser/Ser. As for APE1 Asp148Glu, heterozygous subjects showed significantly higher urinary 8-OHdG concentrations than did those homozygous for Asp/Asp. Moreover, global ethnic comparison of the allelic frequencies of the 4SNPs was performed in 10 population and previous reported data. The mutant allele frequencies of hOGG1 Ser326Cys in the Asian populations were higher than those in the African and Caucasian populations. As for APE1 Asp148Glu, Caucasians showed higher mutant frequencies than those shown by African and Asian populations. Among Asian populations, the Bangladeshi population showed relatively higher mutant allele frequencies of the APE1 Asp148Glu polymorphism. This study is the first to demonstrate the existence of genetic heterogeneity in a worldwide distribution of SNPs (hOGG1 Ser326Cys, APE1 Asp148Glu, XRCC1 Arg280His, and XRCC1 Arg399Gln) in the BER genes.     

Genetic polymorphisms in hMTH1, hOGG1 and hMYH and risk of chronic benzene poisoning in a Chinese occupational population

Oxidative damage to DNA induced by benzene is an important mechanism of its genotoxicity, which leads to chronic benzene poisoning (CBP). Therefore, genetic variation in DNA repair genes may contribute to susceptibility to CBP in the exposed population. We hypothesized that single nucleotide polymorphisms (SNPs) in hMTH1, hOGG1 and hMYH genes are associated with risk of CBP. We genotyped SNPs at codon 83 of hMTH1, codon 326 of hOGG1, and codon 324 of hMYH in 152 CBP patients and 152 healthy workers occupationally exposed to benzene without poisoning manifestations. The genotypes were determined by polymerase chain reaction-restrained fragment length polymorphism (PCR-RFLP) technique. There were 2.51-fold [adjusted odds ratio (OR_adj), 2.51; 95% CI, 1.14-5.49; P = 0.02] and 2.49-fold (OR_adj, 2.49; 95% CI: 1.52-4.07; P < 0.01) increased risk of CBP for individuals carrying genotypes of hMTH1 83Val/Met + Met/Met and hOGG1 326Cys/Cys, respectively. Compared with individuals carrying genotypes of hOGG1 326Cys/Cys and hMYH 324His/His at the same time, there was a 0.33-fold (OR_adj, 0.33; 95% CI: 0.15-0.72; P < 0.05) decreased risk of CBP for those with genotypes of hOGG1 326Ser/Cys + Ser/Ser and hMYH 324His/Gln + Gln/Gln. In the smoking group, there was a 0.15-fold (OR_adj, 0.15; 95% CI, 0.03-0.68; P = 0.01) decreased risk of CBP for subjects carrying genotypes of hMYH 324His/Gln + Gln/Gln compared with those of genotype of hMYH 324His/His. Therefore, our results suggested that polymorphisms at codons 83 of hMTH1 and codon 326 of hOGG1 might contribute to CBP in a Chinese occupational population.     

Rapid detection of the hOGG1 Ser326Cys polymorphism using LightCycler technology

Human 8-oxoguanine DNA glycosylase 1 (hOGG1) plays an important role in the repair of 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodGuo), one of the major constituents in DNA damage. A recent in vitro study showed that the hOGG1 326Cys polymorphism (rs1052133) exhibits reduced 8-oxodGuo repair activity. This study aimed to develop a LightCycler (LC) assay to analyze the C>G polymorphism (Ser326Cys) in exon 7 of the hOGG1 gene followed by validation of the method using DNA samples from 260 polycyclic aromatic hydrocarbons(PAH)-exposed workers with known 8-oxodGuo DNA-adduct values measured by HPLC. Twenty DNA samples were analyzed by a PCR-RFLP analysis with Fnu4H I to generate control DNA. LC melting curve analyses of the hOGG1 exon 7 PCR product were characteristic of the probes hybridized to the non-mutated Ser-type (CC) at 65 degrees C, or to the Cys mutant (GG) at 59 degrees C. The distribution in the population of PAH-exposed workers (N=260) was 58% (CC), 38%(CG), and 4% (GG). The minor G allele displayed a frequency of 23 %. The distribution of 8-oxodGuo adducts for the Ser326Cys variants of hOGG1 revealed geometric means (GM) of 5.83 (CC), 5.27 (CG), and 6.53 (GG) 8-oxodGuo adducts/10(6)dGuo. Corresponding GM values of current smokers were 5.7 (CC), 5.6 (CG) and 7.0 (GG) 8-oxodGuo adducts/10(6) dGuo. The analysis of the Ser326Cys polymorphism in 260 DNA samples with this new LC assay revealed that this method is reliable for high throughput analysis of this key polymorphism in the hOGG1 gene.