PON1基因多态性与乳腺癌的遗传易感性研究.pdf

广西医科大学学报JOURNAL OF GUANGXI MEDICAL UNIVERSITY2023Jul.40（7）PON1基因多态性与乳腺癌的遗传易感性研究*陈坤1，王晓刚2，黄志强1（1.广西壮族自治区桂东人民医院乳腺甲状腺外科，梧州543001；2.广西壮族自治区桂东人民医院输血科，梧州543001）摘要目的：探讨对氧磷酶1基因（PON1基因）rs854560、rs757158两位点与广西地区女性人群乳腺癌（BC）的遗传易感性关系。方法：选取广西地区64例无血缘关系并排除癌症家族史的女性健康体检者（对照组）、60例排除其他肿瘤和遗传病的女性BC患者入组（BC组）。所有参与者清晨空腹采集2 mL EDTA-K2抗凝静脉血样本，从两组的全血样本中提取全基因组DNA。检测两组的PON1基因rs854560 和rs757158 SNP位点。比较两组间的PON1基因rs854560 和rs757158 两位点基因型及等位基因分布频率的差异性，通过校正年龄、吸烟史、饮酒史、民族因素的logistic回归分析筛选独立的易感与保护基因型和等位基因，用比值比（OR）、95%可信区间（95%CI）表示患病的相对风险性，并运用SHEsis在线软件对各组中PON1两个SNP位点进行单倍型构建分析。结果：与对照组比较，BC组年龄、饮酒、吸烟均有显著差异（P0.05），而体质量指数、民族构成比均无显著差异（P0.05）。PON1基因rs854560和rs757158基因型和等位基因在两组间分布频率比较差异均无统计学意义（P0.05），PON1基因 rs854560 和 rs757158 基因型和等位基因与BC无相关性（P0.05），且构建的单倍型AT增加1.662倍BC患病风险（P0.05）。结论：年龄、饮酒、吸烟是BC的危险因素，PON1基因rs854560、rs757158两位点与广西地区女性人群BC及各分子分型之间的患病风险均无关联，PON1基因rs854560和rs757158两位点构建的单倍体AT与BC的发病风险有关，单倍体AT增加BC的患病风险，单倍体AT可能是导致广西地区女性人群BC患病风险的遗传危险因素。关键词乳腺癌；对氧磷酶1；基因多态性；单倍体中图分类号：R394.3；R737.9文献标志码：A文章编号：1005-930X（2023）07-1153-07DOI：10.16190/ki.45-1211/r.2023.07.011Reseach on polymorphisms of PON1 gene and susceptibility to breast cancerChen Kun1,Wang Xiaogang2,Huang Zhiqiang1.(1.Department of Breast and Thyroid Surgery,Guidong People sHospital of Guangxi Zhuang Autonomous Region,Wuzhou 543001,China;2.Department of Blood Transfusion,Guidong People s Hospital of Guangxi ZhuangAutonomous Region,Wuzhou 543001,China)AbstractObjective:To explore the correlation between rs854560 and rs757158 in paraoxophosphate 1(PON1)gene and the genetic susceptibility to breast cancer(BC)in the female population in Guangxi.Methods:A total of 64 unrelated female healthy physical examination subjects without a family history of cancer(controlgroup)and 60 female BC patients excluding other tumors and genetic diseases(BC group)from Guangxi were en-rolled.2 mL EDTA-K2anticoagulant venous blood samples of all participants based on morning fasting were col-lected,and whole-genome DNA was extracted from whole blood samples of both groups.Single nucleotide poly-morphisms(SNPs)of rs854560 and rs757158 in PON1 gene were detected in 64 healthy controls and 60 BC pa-tients.The differences of genotypes and allele distribution frequency of rs854560 and rs757158 in PON1 gene be-tween the two groups were compared.Independent susceptibility and protection genotypes and alleles werescreened by logistic regression analysis by adjusting for age,smoking history,alcohol consumption history,aswell as ethnicity.The relative risk of the disease was expressed by odds ratio(OR)and 95%confidence interval(95%CI).The haplotype construction analysis of two SNP sites in PON1 in each group was performed by SHE-sis online software.Results:Significant differenceswere identified in age,alcohol and smoking con-sumption between the control group and the BCgroup(P0.05),whereas no significant differences*基金项目：广西壮族自治区卫生健康委员会自筹经费课题项目（No.Z2016280）通信作者，E-mail：，Tel：13977497113收稿日期：2022-12-15 1153广西医科大学学报2023 Jul.40（7）were reported in BMI and ethnic composition ratio(P0.05).No statistically significant differences were ob-served in the distribution frequency of genotypes and alleles of rs854560 and rs757158 in PON1 gene betweenthe two groups(P0.05).The genotypes and alleles of rs854560 and rs757158 in PON1 gene were not associat-ed with BC(P0.05),and the constructed haplotype AT had a 1.662-fold increase in the risk of BC(P0.05).Conclusion:Age,drinking and smoking consumption are the risk factors for BC.The two sites of rs854560 andrs757158 in PON1 gene are not associated with the risk of BC and other molecular subtypes in female popula-tion in Guangxi and the constructed haplotype AT of rs854560 and rs757158 in PON1 gene is associated with therisk of BC.Haplotype AT increases the risk of BC,and haplotype AT may be a genetic risk factor for BC in the fe-male population in Guangxi.Keywordsbreast cancer;paraoxonase 1;gene polymorphism;haplotype乳腺癌（breast cancer，BC）是全球妇女最常见的恶性肿瘤1-2，2015年中国总计有近240万例BC新发病例，其中男性约4.4万例3-4。2018年BC的发病率居我国女性恶性肿瘤之首，成为我国女性癌症死亡的主要原因之一5。BC的发病机制复杂，目前尚不清楚。尽管近年来BC在预防和治疗方面取得了巨大的进步，但它仍然给全球广大女性群体的生理和心理健康带来严重威胁6。对氧磷酶1（paraox-onase 1，PON1）基因中rs854560多态性影响PON1酶活性，从而可能增加宿主对各种疾病的易感性7。一些研究结果表明，PON1变异可能对肝细胞癌8、卵巢癌9、BC10以及前列腺癌11有密切的关系。目前PON1基因rs854560和rs757158多态性与广西地区人群BC的相关性研究鲜有报道，本研究旨在探讨PON1基因多态性（rs854560和rs757158）与广西地区女性人群BC发病风险的关系。1资料和方法1.1研究对象选择2017年1月至2018年12月在广西壮族自治区桂东人民医院门诊、住院的60例BC患者（BC组），同期选取本院体检中心的64例无血缘关系并排除癌症家族史的健康女性作为对照组，所有研究对象均来自广西梧州、贺州、玉林、贵港、百色、南宁地区女性人群，病例纳入标准：（1）年龄18岁；（2）首次经病理学检查确诊为BC；（3）未经抗肿瘤药物治疗者（包含化学治疗、内分泌治疗、靶向治疗及免疫治疗）；（4）所有研究对象间无血缘关系；（5）精神状态正常，能顺利配合研究者。排除标准：（1）合并其他肿瘤病史者；（2）合并自身免疫和血液系统疾病者；（3）有遗传病家族史；（4）重要器官功能障碍者。1.2方法1.2.1调查内容本研究获得广西壮族自治区桂东人民医院伦理委员会批准。所有参与者均签署知情同意书，并接受问卷调查与体格检查。问卷调查内容包括年龄、身高、体重、吸烟史、饮酒史、民族等基本情况。按照体重（kg）/身高2（m2）计算体质量指数。1.2.2样本采集所有参与者清晨空腹采集2 mLEDTA-K2抗凝静脉血样本，并提取DNA。1.2.3单核苷酸多态性（SNP）分型采用小测序法（SNaPshot）分型技术平台测定基因型，根据所需检测的位点，设计PCR扩增引物及SNaPshot PCR延伸引物，并由广州擎科生物技术有限公司进行合成。rs854560位点扩增引物序列为：上游：TACAG-AAAGCCTAAGTGAAAGA，下游：TGAGGTGTG-ATAAAGAAATGG。rs757158 位点扩增引物序列为：上 游：CTACACGAGTCAAAAGAACACA，下游：GA