ERAP1基因多态性与丙型肝炎病毒感染及非小细胞肺癌的相关性研究

发布时间：2018-05-23 19:04

本文选题：HCV + NSCLC　；参考：《北京协和医学院》2014年硕士论文

【摘要】：背景：MHCI类抗原呈递基因系统在对抗病毒的有效应答中起着关键作用。内质网氨肽酶1(endoplasmic reticulum aminopeptidase1, ERAP1)是抗原递呈中的重要分子,已有研究报道,ERAP1基因的多态性可影响抗原呈递,引起免疫应答的差异导致与疾病的关联不同。目前的研究集中于自身免疫性疾病,已有多个研究发现ERAP1与自身免疫性疾病的发生发展密切相关。那么,ERAP1基因单核苷酸多态(single nucleotide polymorphism, SNP)是否会影响抗原呈递过程从而与丙型肝炎病毒(hepatitis C virus, HCV)感染、非小细胞性肺癌(non-small cell lung cancer, NSCLC)患病风险相关有待于进一步探讨。此外,云南省由于其独特的地理位置,成为HCV等血液传播疾病传入我国大陆的重要门户,也使其HCV的分布呈现出与其他地区不同的特点。因此,本研究首先开展了ERAP1与HCV感染及NSCLC的相关性研究,在此基础上,结合云南人群HCV感染特点,对不同的HCV基因型与ERAP1基因多态性之间的相关性进行了初步探讨。方法：本研究分为三部分。第一部分以云南地区HCV感染者376名、正常健康对照324名为研究对象,研究ERAP1基因单核苷酸多态性与HCV感染的相关性。采用Taq Man探针基因分型方法检测云南汉族正常人群及HCV感染者的ERAP1基因单核苷酸多态性,进行HCV感染的关联性分析。第二部分以云南地区HCV一般感染人群(general population, GP)为研究对象,首先研究一般感染人群中HCV基因型和亚型的流行状况。采用RT-PCR的方法扩增HCV NS5B片段并测序,对云南一般人群HCV感染者进行HCV基因分型,探索云南一般感染人群HCV流行趋势。在此基础上,结合HCV感染者的ERAP1基因信息,分析不同HCV基因型与ERAP1基因多态性的相关性。第三部分以云南地区非小细胞肺癌患者224例、正常健康对照组207例为研究对象,研究ERAP1基因单核苷酸多态性与非小细胞肺癌的遗传易感性。采用Taq Man探针基因分型方法对ERAP1基因单核苷酸多态性进行分析,探索ERAP1基因单核苷酸多态性与非小细胞肺癌患病风险的相关性。结果：(1)HCV感染组和对照组4个ERAP1基因SNP位点基因型的分布符合HWE平衡,人群具有代表性。其中rs26618基因型和等位基因型频率在病例组和对照组中的差异具有统计学意义(P0.05),病例组和对照组rs27044/rs30187/rs26618/rs26653-CCCG的单倍型频率差异具有统计学意义,P值为0.029(P0.05)。(2)云南地区一般感染人群存在4种HCV基因型(1、2、3和6),7种HCV亚型(1b、2a、3a、3a、6a、6n和6k)。HCV3型是主要流行的基因型,频率为0.484,其次是HCV1型(0.283)、6型(0.133)和2型(0.100)；其中HCV3b亚型(0.292)和1b亚型(0.283)是主要流行的亚型。云南一般感染人群HCV的分布呈现出与其他地区不同的特点：HCV1和3型的频率在云南和重庆、云南和广东具有显著性的差异(P0.05),而在云南和广西的频率分布相似；HCV6型的频率在云南和广西、广东、越南、缅甸地区分布存在显著性差异(P0.05),而云南和重庆的频率分布却相似。另一方面,云南一般感染人群与静脉吸毒人群HCV基因型分布相比较亦存在显著性差异：HCV基因型(1、2和6)及其亚型(1b、2a、6a和6n)在一般感染人群与静脉吸毒人群中的频率有显著性差异(P0.05),经HCV3a和6a构树分析,一般感染人群与静脉吸毒人群的HCV基因型分布呈“交叉”样。ERAP1rs30187位点的基因型频率在不同的HCV基因型(1,2,3和6)和不同的HCV亚型(lb,2a,3a,3b和6n)中有显著性的差异(P0.05)。(3) NSCLC病例组和对照组2个ERAP1基因SNP位点基因型的分布符合HWE平衡,人群具有代表性。ERAPl基因rs26618和rs26653基因型和等位基因频率差异有统计学意义(P0.05)。病例组和对照rs26618/rs26653-CG、TC的单倍型频率差异有统计学意义(P0.05)。而ERAP1基因rs26618、rs26653位点的基因型和等位基因的频率在腺癌和鳞癌、Ⅰ+Ⅱ期和Ⅲ+Ⅳ期非小细胞肺癌患者中的差异不具有统计学意义(P0.05)。结论：(1) ERAP1基因多态性与HCV感染有着显著的易感关联,rs27044/rs30187/rs26618/rs26653-CCCG单倍型可能增加HCV感染的患病风险(OR=1.311,95%CI:1.028-1.671).(2)云南地区一般感染人群HCV基因型和亚型的分布与静脉吸毒人群有着明显的不同,且HCV基因型和亚型正从静脉吸毒人群向一般感染人群传播。不同的HCV基因型、亚型与ERAP1基因多态性有一定的关联。(3) ERAP1基因多态性与NSCLC的发生有关,rs26618/rs26653-CG单倍型可能增加NSCLC的患病风险(OR=1.725,95%CI:1.270-2.343), rs26618/rs26653-TC单倍型对NSCLC可能具有保护性作用(OR=0.668,95%CI:0.510-0.874).
[Abstract]:Background: MHCI antigen presenting gene system plays a key role in the effective response to the virus. Endoplasmic reticulum aminopeptidase 1 (endoplasmic reticulum aminopeptidase1, ERAP1) is an important molecule in antigen presentation. It has been reported that the polymorphism of the ERAP1 gene can affect the antigenic presentation, causing differences in immune response and disease. The current study is focused on autoimmune diseases. Many studies have found that ERAP1 is closely related to the development of autoimmune diseases. Then, the ERAP1 gene single nucleotide polymorphism (single nucleotide polymorphism, SNP) will affect the antigen presentation process and the hepatitis C virus, HCV, and HCV. The risk of infection, non small cell lung cancer (non-small cell lung cancer, NSCLC) remains to be further discussed. In addition, Yunnan province has become an important gateway to the mainland of our country because of its unique geographical location, and the distribution of HCV is different from other regions. Therefore, this study is the first. The correlation between ERAP1 and HCV infection and NSCLC was first carried out. On this basis, the correlation between different HCV genotypes and ERAP1 gene polymorphisms was preliminarily discussed in combination with the characteristics of HCV infection in Yunnan population.
Methods: This study was divided into three parts. In the first part, the correlation between single nucleotide polymorphisms of ERAP1 gene and HCV infection was studied in 376 people with HCV infection in Yunnan and 324 normal healthy controls. The single nucleotide polymorphisms of ERAP1 gene in the normal and HCV infected people of Yunnan were detected by Taq Man probe genotyping. The correlation analysis of HCV infection. In the second part, the HCV general infection population (general population, GP) in Yunnan area was used as the research object. First, the prevalence of HCV genotype and subtype in the general infection population was studied. RT-PCR was used to amplify NS5B fragment and sequence of HCV in the general population of Yunnan, and HCV gene score was carried out for HCV infected persons in general population of Yunnan. On the basis of ERAP1 gene information of people infected with HCV, the correlation between different HCV genotypes and ERAP1 gene polymorphisms was analyzed on the basis of the ERAP1 gene information of the people infected with Yunnan. The third part was the study of 224 cases of non small cell lung cancer in Yunnan and 207 cases of normal healthy control group, and the single nucleotide polymorphisms of ERAP1 gene were studied. The genetic susceptibility to non small cell lung cancer was analyzed by the Taq Man probe genotyping method, and the correlation between the single nucleotide polymorphisms of the ERAP1 gene and the risk of non small cell lung cancer was explored.
Results: (1) the distribution of the SNP loci of the 4 ERAP1 genes in the HCV infection group and the control group conformed to the HWE balance, and the population was representative. The difference between the rs26618 genotype and the allele genotype frequency in the case group and the control group was statistically significant (P0.05), and the frequency of the haplotype of the case group and the rs27044/rs30187/rs26618/rs26653-CCCG group was the same. The difference was statistically significant, the P value was 0.029 (P0.05). (2) there were 4 HCV genotypes (1,2,3 and 6) in the general infection population in Yunnan, and 7 HCV subtypes (1b, 2a, 3a, 3a, 6a, 6N and 6K) were the main genotypes, the frequency was 0.484, followed by 0.283, 6 (0.133) and 2 (0.100). The distribution of HCV in Yunnan is different from other regions: the frequency of HCV1 and type 3 has significant differences in Yunnan and Chongqing, Yunnan and Guangdong (P0.05), while the frequency distribution in Yunnan and Guangxi is similar; the frequency of HCV6 is divided in Yunnan and Guangxi, Guangdong, Vietnam and Burma. There were significant differences in distribution (P0.05), while the frequency distribution in Yunnan and Chongqing was similar. On the other hand, there was a significant difference in the distribution of HCV genotypes between the general infection population of Yunnan and the intravenous drug users: the HCV genotype (1,2 and 6) and its subtypes (1b, 2a, 6a and 6N) had a significant frequency in the general infection population and the intravenous drug users. P0.05, after HCV3a and 6A tree analysis, the genotype frequencies of the HCV genotype distribution of the general infected people and the intravenous drug users were "cross" like.ERAP1rs30187 loci in different HCV genotypes (1,2,3 and 6) and different HCV subtypes (LB, 2a, 3a, 3b, etc.). (3) 2 cases and control groups The distribution of the genotype SNP loci of the RAP1 gene conforms to the HWE balance, and there is a significant difference in the frequency difference between the rs26618 and rs26653 genotypes and the allele frequencies of the representative.ERAPl genes (P0.05). The frequency difference between the case group and the control rs26618/rs26653-CG, TC is statistically significant (P0.05). The frequencies of genotype and allele were not statistically significant in patients with adenocarcinoma and squamous cell carcinoma, stage I, + II and III + IV patients (P0.05).
Conclusion: (1) there is a significant association between ERAP1 gene polymorphism and HCV infection, and rs27044/rs30187/rs26618/rs26653-CCCG haplotype may increase the risk of HCV infection (OR=1.311,95%CI:1.028-1.671). (2) the distribution of HCV genotypes and subtypes in the general infection population in Yunnan region is significantly different from that of intravenous drug users, and the HCV base The type and subtype are spreading from the intravenous drug users to the general infection population. Different HCV genotypes and subtypes are associated with the polymorphism of the ERAP1 gene. (3) the polymorphism of the ERAP1 gene is associated with the occurrence of NSCLC, and the rs26618/rs26653-CG haplotype may increase the risk of NSCLC (OR=1.725,95%CI:1.270-2.343), and rs26618/rs26653-TC haploidy. Type NSCLC may have protective effects on OR=0.668,95%CI:0.510-0.874.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R512.63;R734.2

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