【作者】 吴开松；

【导师】 周新；

【作者基本信息】 武汉大学 ， 内科学， 2010， 博士

【摘要】 背景原发性支气管肺癌(primary bronchogenic carcinoma),简称肺癌(lung cancer),是我国常见的恶性肿瘤之一,发病率和死亡率均较高,并呈逐年上升趋势。肺癌发生的危险因素包括环境和机体遗传因素,以及二者的交互作用。环境因素中吸烟在肺癌发病中的地位已得到多方面证据的支持。IL-1基因常被作为研究基因与环境相互作用的常见基因。IL-1基因位于2q13,包括IL-1α、IL-1β和1个特异性受体拮抗因子IL-1ra。IL-1β是机体免疫炎症反应中的关键因子,已被证实是气道的重要炎症介质,广泛参与气道炎症。IL-1β在启动子-31、-511处存在C-T单核昔酸多态性(SNP),多态性导致IL-1局部生成过量和/或IL-1ra生成减少,与胃癌的发生及多种感染性疾病的的发生和严重程度相关。Zienolddiny S等研究发现IL-1β-31T/T及-511C/C增加挪威人非小细胞肺癌的发病风险。这两个基因的多态性与汉族人非小细胞肺癌发生中是否有某种关联?同时,其效应是否受到环境等因素的影响?本文从分析肺癌的病理与临床入手,探讨IL-1β-31、-511基因多态性和环境暴露因素的交互作用对汉族人非小细胞肺癌发生的影响以及可能的机制。目的分析肺癌的病理与临床之间的联系,总结肺癌的临床特点,为肺癌的早期诊断提供可能的依据；研究位于2号染色体上IL-1β基因多态性与非小细胞肺癌的发病风险的关系；探讨烟、酒与这组非小细胞肺癌易感基因多态性的交互作用及可能的发病机制。方法运用SPSS13.0软件对我院2002年1月-2008年12年确诊的329例肺癌患者肺癌的病理类型与年龄、性别、部位及临床表现等的关系进行逐一分析。采用病例对照的方法研究基因、环境与肺癌发生的关系。采用健康问卷的方式收集肺癌患者和健康对照者烟酒、饮食等基本信息。健康问卷的内容主要有：人口学基本数据、吸烟情况、饮酒习惯、家族史、结核病病史等等；IL-1β-31T/T及-511C/C位点基因多态性的检测,采用多聚酶链反应-限制性片段长度多态性(polymerase chain reaction—restriction fragment length polymorphism, PCR-RFLP)方法。通过基因克隆、载体表达技术研究IL-1β启动子-31C/T多态性对IL-1β表达影响。统计分析：采用SHEsis软件估计单倍型频率,并分析人群中IL-1β-31T/T及-511C/C基因间是否存在连锁不平衡；应用logistic回归模型分析多因素与非小细胞肺癌发生的关系,估计相对危险度,即优势比(odds ratio, OR)。结果入组329例肺癌患者,年龄58.73±9.85岁,男227例,女102例。其中腺癌107例(32.5%))、鳞癌145例(44.1%)、腺鳞癌49例(15.0%)、小细胞癌28例(8.5%)。性别构成比,男性明显高于女性,特别是在鳞癌患者中。各病理类型以中老年为主。鳞癌和小细胞癌以中央型多见,腺癌以周围型为主。临床就诊患者多为Ⅲ期、Ⅳ期肺癌。在调整了年龄、性别等混杂因素的条件下,logistic回归分析显示研究人群中非小细胞肺癌的传统危险因素仍是吸烟(OR=10.13,95%CI：4.58-22.42)。IL-1-31基因T/C基因型在人群中常见,T等位基因与非小细胞肺癌的发生密切相关(OR=3.247,95%CI:1.794-5.878)。IL-1-511基因T/C基因型在人群中常见,本组病例未发现该位点多态性与非小细胞肺癌的发病风险相关。IL-1-31基因与IL-1-511基因存在连锁不平衡(D=0.827,r2=0.785),IL-1β-31T/-511C单倍体型能显著增加非小细胞肺癌的发病风险(OR=1.531,95%CI：1.163～2.814)。IL-1基因与吸烟间呈现显著的交互作用,其对发生非小细胞肺癌的危险性在各基因型与吸烟的量有关,其中IL-1β-31 T/T型-重度吸烟(15.246)风险最高。IL-1基因与饮酒间也呈现显著的交互作用：轻度饮酒未发现增加非小细胞肺癌的发病风险,但重度饮酒能增加IL-1β-31 T/T型(OR＝1.913)和IL-1β-511C/C型(OR=5.022)个体的发病风险。IL-1β启动子-31T等位基因能增加IL-1β的表达。结论1.在本研究人群中,吸烟仍是非小细胞肺癌发生的危险因素。2.位于2号染色体q13-q14的IL-1-31和IL-1-511两基因存在显著的连锁不平衡,IL-1-31T/C多态性与非小细胞肺癌发生密切相关。3.IL-1-31T/T和IL-1-511C/C基因等位基因携带者吸烟和重度饮酒显著增加非小细胞肺癌发病风险。4.IL-1β启动子-31T等位基因能增加IL-1β的表达。更多还原

【Abstract】 BackgroundPrimary bronchogenic carcinoma also called lung cancer, is one of the most common malignant tumors in China, the incidence and mortality rates are high and increasing year by year. The risk factor of lung cancer includes environmental and genetic factors, and their interactions. Among environmental factors, the status of smoking in the pathogenesis of lung cancer has been supported by various evidences. IL-1 gene was used as one common candidate gene for the study of interaction of environment and genes.IL-1 gene lies at the long arm of chromosome 2(2q13), including IL-1α,IL-1βand a specific receptor agonist IL-1ra. IL-1βis a key factor in the inflammatory response by the body, which has been proved to be an important mediator of inflammatory in the airway and was widely involved in airway inflammation.Zienolddiny S and other study found that IL-1β-31T/T and-511C/C increase the incidence of non-small cell lung cancer in Norwegian. Is there any association between polymorphisms of this two genes and the incidence of non-small cell lung cancer in Han people?At the same time, whether its effects are influenced by environmental and other factors? This article, basing on the analysis of pathology features and clinical presentation of lung cancer, studys the gene polymorphism of IL-1β-31,-511 and their interactions with environmental factors in occurrence of non-small cell lung cancer in Hans as well as its relevant mechanisms.ObjectiveTo analyze the relationship between pathological and clinical presentation of lung cancer, summarize the clinical characteristics of lung cancer and provide credible basis for the early diagnosis of lung cancer.To investigate the association of gene polymorphisms on chromosome 2 with the risks of non-small cell lung cancer. To find the interplay between the behavioral factors (such as drinking and smoking) with genetic polymorphisms in non-small cell lung cancer and its possible mechanisms.Methods329 lung cancer patients were diagnosed in ZhongNan hospital between 2002.1-2008.12.Their pathology type of cancer、age、gender, position of the cancer and clinical manifestations were analyzed one by one by the SPSS 13.0 software.A case-matched study was carried out to investigate the relations among gene, environment and the occurrence of lung cancer. Use the healthy questionnaire way to collect basic information of lung cancer patients and healthy collators about smoking and drinking, diet and so on. The healthy questionnaire mainly includes:Demography materials, smoking and drinking, diet custom, family history, formerly medical history and so on. Genetic polymorphisms of IL-1β-31T/T and-the 511C/C position were analyzed by polymerase chain reaction-restrictive fragment length polymorphism (PCR-RFLP).The influence of IL-1βpromoter-31C/T’s polymorphism on the IL-1βexpression was studied by gene cloning, carrier expression engineering.Linkage disequilibrium between the IL-1-31 and IL-1-511 was tested by estimation of haplotype frequencies performed by the SHEsis program. And OR of susceptibility of the genes to non-small cell lung cancer was analyzed by using multivariate logistic regression.ResultsThere are 329 (male 227, female 102) patients of lung cancer, aged 58.73±9.85 years old. Their pathologic diagnoses are as follows:adenocarci-noma 107 cases (32.5%), squamours cancer 145 cases (44.1%), adenosquamous carcinoma 49 cases (15.0%), small cell carcinoma 28 cases (8.5%). There are more male than female, especially in adenocarci-noma. The main pathological type is middle aged and elderly people. The central type was most common in squamours cancer and small cell carcinoma, while peripheral in adenocarci-noma. Most of them were of stageⅢorⅣof lung cancer.From the stepwise logistic regression analyses:smoking emerged as significant risk factor for non-small cell lung cancer (OR=10.13,95%CI:4.58-22.42) For IL-1-31 gene, the T/C genotypes represent the most common in the population. (OR=3.247,95%CI:1.794-5.878)For IL-1-511 gene, the T/C genotypes represent the most common in the population.The polymorphism of this gene is not related with non-small cell lung cancer.A linkage disequilibrium was revealed between IL-1-31 gene and IL-1-511 gene. (D =0.827, r2=0.785) Haplotype of IL-1β-31T/-511C significantly increased the incidence of non-small cell lung cancer. (OR=1.531,95%CI:1.163-2.814)A significant interaction showed between IL-1 gene and smoking, the incidence of non-small cell lung cancer on the risk of the genotype was related to smoking volume, among which IL-1β-31 T/T type-heavy smokers (15.246) have the highest risk.IL-1 gene also showed significant interaction with alcohol:mild alcohol consumption was not found to increase the risk of non-small cell lung cancer, but heavy drinking can increased the incidence of the individual,who carrier IL-1β-31 T/T type (OR=1.913) and IL-1β-511C/C type (OR=5.022).IL-1βpromoter-31T allele increased the expression of IL-1β.Conclusion1. In this study, smoking is still a risk factor of non-small cell lung cancer.2. Q13-q14 of the IL-1-31 and IL-1-511 genes, located on chromosome 2, had significant linkage disequilibrium. Polymorphism of IL-1-31T/C is highly related to non-small cell lung cancer.3. People who carriers IL-1-31T/T and IL-1-511C/C allele and smokes and have heavy alcohol consumption significantly increased the risk of non-small cell lung cancer.4. IL-1βpromoter-31T allele increases the expression of IL-1β.更多还原