摘要
目的:探寻幽门螺杆菌(Helicobacter pylori,Hp)铁摄取调节蛋白(ferric uptake regulator,Fur)基因单核苷酸多态性(single nucleotide polymorphism,SNP)和进化分型与胃癌的相关性.方法:选取2011-2018年青岛市市立医院保存的150株Hp(胃癌来源59株和胃炎来源91株),运用聚合酶链式反应(polymerase chain reaction,PCR)方法扩增fur基因,并进行一代测序及SNP分析.通过NCBI数据库下载226株东亚亚群Hp 菌株fur基因序列,应用MEGA 5.0软件分析SNP并构建fur基因Neighbour-Joining系统进化树,建立进化分型.结果:98.7％(148/150)Hp菌株fur 基因PCR扩增阳性.序列分析发现fur基因351位点存在碱基A→G的同义SNP(SNPA351G),胃癌来源的菌株中G等位基因的变异频率明显高于胃炎来源的菌株,差异有统计学意义(x2=5.161,P=0.023);携带该等位基因的菌株发生胃癌风险明显升高(OR=2.4).在东亚Hp fur基因的Neighbour-Joining系统进化树中,依据进化距离将东亚Hp菌株分为Ⅰ型和Ⅱ型两个亚型,fur基因进化Ⅰ型中的胃癌来源Hp菌株比例明显高于Ⅱ型,差异有统计学意义(x2=41.8,P=9.9×10-11);感染furⅠ型Hp的患者发生胃癌的风险显著升高(OR=4.7).结论:携带fur SNPA351G的Hp菌株导致胃癌发生风险显著升高,fur基因进化 Ⅰ型与胃癌发生风险具有一定相关性.
Abstract
Objective:To explore the single nucleotide polymorphism(SNP)and evolutionary characteristics of the ferric uptake regulator(Fur)gene of Helicobacter pylori(Hp)associated with gastric cancer.Methods:150 Hp strains(59 strains from patients with gastric cancer and 91 strains from gastritis patients)were selected which were stored in Qingdao Municipal Hospital from 2011-2018.The fur gene was amplified by polymerase chain reaction(PCR),then the generation sequencing and SNP analysis were proceeded.226 Hp fur sequences of East Asia subgroup were downloaded from the NCBI database.SNPs were analyzed and a Neighbour-Joining phylogenetic tree was constructed to establish evolutionary typing using MEGA 5.0 software.Results:98.7％(148/150)of the Hp strains were positive for fur gene using PCR amplification.A synonymous SNP of base A→G(SNP A351G)existed at site 351 of the fur gene.The variation frequency of G allele had a significantly higher in gastric cancer strains than that of gastritis strains(x2=5.161,P=0.023).The risk of gastric cancer significantly increased in strains carrying this allele(OR=2.4).Neighbour-Joining phylogenetic tree was separated into two phylogenetic subgroups,type Ⅰ and type Ⅱ based on the sequence of the Hp East Asia fur gene.The prevalence of Hp strains of patients with gastric cancer was predominantly higher in the evolutionary type Ⅰ subgroup(x2=41.8,P=9.9×10-11).The risk of gastric cancer was significantly higher with fur gene evolutionary type Ⅰ(OR=4.7).Conclusion:Hp carrying fur SNP A351G caused a significantly higher risk of gastric cancer,and fur gene evolutionary type Ⅰ had an association with the risk of gastric carcinogenesis.
NETL
NSTL
维普
万方数据