【作者】 武治国；

【导师】 陈明清； 董坚； 李文亮；

【作者基本信息】 昆明医学院 ， 外科学， 2010， 硕士

【摘要】 目的：通过对云南省家族性腺瘤性息肉病(FAP)家系的收集,抢救和保存云南省FAP家系遗传资源,为FAP预防、诊治及病因的研究提供遗传性标本。同时,对FAP相关致病基因APC基因进行分析,探讨云南省FAP相关致病基因突变特点,初步建立符合云南省FAP家系的筛查方案。方法：对昆明医学院第一附属医院、云南省肿瘤医院、红河州和大理州等多家医院近10年的结直肠癌患者的病案调查,筛选FAP患者,以及这些医院近3年门诊、肠镜室、相关科室新发FAP患者的收集,通过电话联系患者并登门随访,经知情同意后,由其介绍和引导其直系和旁系各级亲属进行遗传学咨询,并以先证者为中心绘制家系图谱,采集患者及其家系成员外周静脉血,及提取手术患者离体后组织标本(包括肿瘤、瘤旁、远端正常组织)、石蜡包埋切片,分别提取DNA,利用已建立的云南省遗传性大肠癌标本库进行标本保存和信息管理。应用聚合酶链反应(PCR)方法对FAP家系先证者DNA进行APC基因扩增,DNA直接测序法进行APC基因全编码区测序,应用Mutation Surveyor软件分析测序结果,参考美国国家生物技术信息中心(NCBI)网站上APC基因序列,进行序列比对,确定突变位点,再根据突变位点筛查其家系高危亲属。结果：共收集10个FAP家系,其中包括1个白族家系,2个彝族家系和7个汉族家系,并采集家系中主要成员血液标本96份,组织标本4份,蜡块组织7份,并提取DNA,保存于云南省遗传性大肠癌标本库。10个FAP家系先证者中,1个汉族家系先证者检出APC基因致病突变(10%),此突变存在于APC基因第15外显子c.3587C>A(S1196X);随后对该家系另外8名成员进行该突变位点筛查,其中7人有此突变；其余9个FAP家系未查出APC基因有意义的突变。结论：通过对云南省FAP家系的收集,抢救和保存云南省FAP家系宝贵的遗传资源,为后期FAP预防诊治及病因的研究提供现代分子肿瘤学要求的遗传性标本,提高云南省遗传性大肠癌标本库的应用价值。首次在云南省开展FAP家系APC基因突变分析,并发现国内尚未见报道的APC基因15号外显子c.3587C>A(S1196X)无义突变。云南省FAP患者APC基因致病突变发生率(10%)明显低于国内外报道,未检出APC基因致病突变的FAP患者可能存在其他发病的机制。针对目前APC基因低突变结果,初步建立符合云南省FAP家系的基因和肠镜两套筛查方案。更多还原

【Abstract】 Objective:To rescue and preserve the genetic resources of familial adenomatous polyposis (FAP) by collecting FAP pedigrees in Yunnan Province,and to provide genetic samples for prevention, treatment and etiology research of FAP. Meanwhile, to analysis the APC gene that predispose the disease susceptibility in patients with FAP from Yunnan Province and explore the mutation features of disease gene FAP-related and initial establish suitable program of screening FAP pedigree in Yunnan Province.Methods:Screening FAP patients through investigating the cases of colorectal cancer patients in the past 10 years and collecting new FAP patients of the endoscopy room, out-patients and the relevant departments nearly 3 years from the First Affiliated Hospital of Kunming Medical College, Yunnan Province Cancer Hospital, Red River State Hospital and Dali State Hospita. Using patients contacted by phone and door follow-up as index patients, the pedigree and all lineal and collateral relatives were interviewed. Peripheral venous blood samples of individuals were collected from these families. Collecting tissue specimens (including tumors, normal tissues beside or outside tumors) and paraffin-embedded sections of operative patients. These specimens were preserved in the Yunnan Province of hereditary colorectal cancer tissure bank. The whole coding region of APC gene of the FAP family probands were screened for germline mutations by using polymerase chain reaction (PCR) and DNA direct sequencing, and Mutation Surveyor software analyze sequencing results. Reference the APC gene sequence on the NCBI website to determine the sites of mutation and according to the sites of mutation screening high-risk relatives of their family.Results:Total of 10 FAP pedigrees were collected, including a Bai pedigree,2 Yi pedigrees and 7 Han pedigrees. There are 96 blood specimens,4 tissue specimens and 7 paraffin-embedded sections.DNA of all family members are extracted in reserve. Among the 10 probands subjected to genetic analysis of the APC gene, mutation was detected in 1 han proband (10%), which is the C to A transversion resulting in a stop codon at codon 1196 within exon15(c.3587C＞A,S1196X).Subsequently, another 8 members in APC gene mutaition family,7 of them have this mutation. The remaining 9 FAP pedigrees without identified significant mutation of APC gene.Conclusions:Through the collection of FAP family in Yunnan Province, rescue and preserve valuable genetic resources of FAP pedigrees, in order to provid the genetic samples of requirements of modern molecular oncology for future treatment, prevention and etiology research of FAP, and improve the value of the Yunnan Province of hereditary colorectal cancer tissure bank. We first analyze the APC gene mutations of FAP pedigree in Yunnan Province and found nonsense mutation has not been reported that the APC gene exon 15 c.3587C＞A(S1196X) in China. Among patients with FAP in Yunnan Province, the incidence of APC gene mutations was significantly lower than other areas at home and abroad is reported. others pathogenesis may exist in FAP patients without APC gene mutation. Aiming at results of APC gene mutations with low, establish two sets of suitable program of screening FAP pedigree in Yunnan Province.更多还原