【作者】 李晓燕；

【导师】 王贵齐； 高燕宁； 张开泰；

【作者基本信息】 中国协和医科大学 ， 肿瘤学， 2010， 硕士

【摘要】 腺癌发病率逐年提高。周围型肺腺癌为临床诊断的一个难题,目前缺乏有效易行的鉴别诊断手段。有关区域性癌化现象在气道上皮细胞中的研究表明,形态正常的气道上皮细胞分子水平的改变可以用来辅助诊断肺癌,但是针对不吸烟肺腺癌患者的相关研究很少。本课题在实验室已有的稳定的微阵列比较基因组杂交技术(array comparative genome hybridization Array-CGH)平台基础上,研究不吸烟周围型肺腺癌患者与良性周围型疾病患者的支气管上皮细胞基因组DNA拷贝数异常的变化情况,以期获得NSPLAd患者及PBPD患者支气管上皮细胞基因组DNA拷贝数改变谱型；进而揭示NSPLAd患者支气管上皮的特征性改变的基因或DNA片段,并以这些DNA片段的组合来进行NSPLAd患者的鉴别诊断,以指导正确治疗。首先,应用Agilent Human Genomic CGH 44B芯片,对16例患者(11例NSPLAd,5例PBPD)的32个部位的支气管上皮细胞刷片(每位患者包括健侧叶段支气管上皮细胞-1s及患侧主支气管上皮细胞-2s)基因组DNA进行了Array CGH检测。继而,采用Agilent CGH Analytics 4.0和GeneSpring GX 11.0.1分析软件分析实验数据。结果显示：(1)11个NSPLAd患者支气管上皮细胞基因组DNA拷贝数发生异常的区域较一致,1s发生频率大于45%片段有15个,2s发生频率大于50%的DNA片段有10个。(2) NSPLAd患者与PBPD患者对应的支气管上皮相比,基因组DNA拷贝数变异的区域广,范围大,发生频率高。(3)经过对11例患者2s处Array CGH实验数据的生物信息学分析,初步筛选出34个能用于鉴别诊断NSPLAd患者的DNA片段组合。综上,本研究表明,NSPLAd患者形态正常的支气管上皮中存在一系列基因组DNA拷贝数变异,2s处基因组DNA拷贝数变异可能用于NSPLAd的辅助诊断,有望通过进一步扩大样本量的深入研究建立起能够用于NSPLAd鉴别诊断的分子模型。更多还原

【Abstract】 Diagnosis of Non-smoking Peripheral Lung Adenocarcinoma by Bronchial Epithelial Cell Genomic DNA Copy Number VariationLung Adenocarcinomas incidence are increasing。Peripheral lung Adenocarcinoma (PLAd) is difficult pclinically。Effective and easy means to differentiate PLAd is not always available. Field cancerization exists in airway epithelial cells, and a series of molecular changes in the airway epithelial cells showed useful in the differential diagnosis of lung cancer. Such researches in non-smoking lung cancer patients are rare. In this study, array comparative genomic hybridization (Array-CGH) was performed on the bronchial epitherial cells of non-smoking patients with peripheral lung adenocarcinoma (NSPLAd) and patients with benign peripheral disease (PBPD),to identify the DNA copy number variation(CNV) profiles that specially related with NSPLAd and PBPD patients, respectively. From these, a set of gain or loss genes/regions could be identified to distinguish the two disease.The 32 bronchial epithelial cells (including the contralateral bronchial epithelial cells-1s and ipsilateral primary bronchial epithelial cell-2s) derived from 16patients (11 NSPLAd and 5 PBPD)were screened by Agilent Human Genomic CGH 44B array.With analyses by Agilent CGH Analytics 4.0 and GeneSpring GX 11.0.1, the genome-wide CNV profiles of bronchial epithelial cells were obtained.11 NSPLAds showed good accordance in regions of CNV. There were 15 copy number alterations with a frequency of more than 45% in 1s and 10 copy number alterations with a frequency of more than 50% in the NSPLAd. NSPLAd patients showed a much larger scale, much higher frequence in CNV. than PBPD. Bioinformatical analysis was performed with the array CGH data of the 32cases, and 34 genes that could differentiate NSPLAd from PBPD were screened out.In conclusion, data obtained from a reliable array CGH analysis indicated that there were a series of differences in the genome-wide CNV profiles in the bronchial epithelial cells of the NSPLAd from PBPD. It was promised to build up a molecular model to distinguish NSPLAd from PBPD with bronchial epithelial cells, by further invegestion with an enlarged sample size.更多还原