【作者】 路军；

【导师】 吕宁； 张开泰； 薛丽燕；

【作者基本信息】 中国协和医科大学 ， 病理学与病理生理学， 2010， 博士

【摘要】 背景：食管鳞癌是我国癌症疾患的主要病种之一,早期诊断早期治疗具有重要意义。随着肿瘤诊治理念的进步,近年对早期食管鳞癌探索实施局部粘膜切除术等新方案,达到在避免过度治疗的前提下根治肿瘤的目的。新方案所面临的关键问题是能否正确选择适应症。目前无论是对活检标本病理学检查和其他影像学检查均难以判断肿瘤的进展和转移倾向。肿瘤进展转移是一个极其复杂的多基因参与,多基因调控的过程,因此从分子水平上检测与肿瘤相关遗传学的改变,寻找和揭示与早期食管鳞癌进展和转移相关的分子生物学标志物,从而提供判断或预测肿瘤进展转移的分子水平依据是本项目研究探索的主要目标。本研究采用微阵列比较基因组杂交(Array-based Comparative genomic hybridization, Array-CGH)及TaqMan Human MicroRNA芯片技术,试图从早期食管鳞癌组织的“DNA片段拷贝数变化谱型”及"microRNA表达谱”中筛选出有鉴别意义的扩增(或缺失)基因／染色体区段和microRNA,以此作为早期食管鳞癌进展转移潜能相关的标志物,从而为选择正确的治疗方案提供科学依据。方法：第一部分：采用Array-CGH技术检测24例T1N0期食管鳞癌组织中全基因组染色体DNA拷贝数改变,研究病例中11例患者术后40月内死亡(死亡组),13例患者术后生存80月以上(生存组)。第二部分：采用TaqMan Human MicroRNA Arrays技术分析第一部分所用两组病例中的16例microRNA表达情况：生存组和死亡组各选取前8例。通过以上技术分析早期食管鳞癌组织全基因组染色体拷贝数改变及microRNA表达与肿瘤进展及转移等预后关系。结果：1、两组样品中共有多条染色体发生变化,其中最常见的染色体基因组扩增发生在1 q、3 q、5p、6p、7q／p、8q／p、9q、11 q、12p、14q、16p、17 q／p、18p、19 q／p、20 q／p和22q染色体区段上,在染色体3p、4q／p、5q、6p、9p、11 q、13 q、14q、18q、19q、21q中常见染色体基因组缺失。与死亡组相比较,生存组更易出现包括染色体1 q、3q、5p、7q／p、8q／p、9q、11 q、12p、14q、16p、17q、19q、20q和22q扩增和染色体4q／p、5q和13q缺失。两组差异性小片段DNA拷贝数改变主要为1q21.3-23.3、3q21.1-24、3q26.1-26.33、5p13.1-15.31、7p22、7q11.23、8p11.2、8q22.1-24.3、11q12.1-13.5、12p12.1-13.33、14q23.1、16p12.1-13.3、17q25.1、19q13.12、20q11.21-13.33、22q12.1-13.2扩增及4p、4q31-35、5q22.3、13q12.21-12.2缺失。进一步分析发现,109个基因及探针可将死亡组与生存组区分开来(p<0.01)。2、两组样品中多个microRNA表达发生改变,经过组间对比,其中四个microRNA表达具有统计学意义(p<0.05),包括：miR-574-3p、MiR-19b、miR-31及miR-28-3p。结论：在早期食管鳞癌显著不同生存期患者的染色体DNA拷贝数变化及microRNA表达变化中,存在特定的染色体区段或基因及microRNA,可能决定肿瘤进展转移潜能。所筛选出的基因组DNA拷贝数改变及相关基因和microRNA有可能成为早期食管鳞癌预测进展转移和预后的分子标志物,从而为早期食管鳞癌患者选择正确的治疗方案提供科学依据。更多还原

【Abstract】 Background:Esophageal squamous cell carcinoma (ESCC), especially in northern China, is one of the common fatal cancers. ESCC is an aggressive cancer with poor prognosis. Most patients present with advanced stage disease when they were diagnosed, so they lost the best time for therapy. Early detection and therapay remains the best way for a cure. With the improvement of tumor therapy, the recommended therapy of early ESCC is endoscopic mucosal resection (EMR). The key point for EMR is to estimate the condition of infiltration of lesions and metastasis exactly, and the metastasis condition of lymph node directly determines the treatment modes. However, there are no effective examinations to evaluate the progression and metastasis of tumors. The most important reason for different tumors with different biological behaviours is that each tumor has its characteristic genetic alterations. Unfortunately there are no specific makers of the genetic alterations for progression and metastasis of tumors, so it would significantly be rational strategies to systemically analyze cancer tissues on molecular level and to find specific molecular markers for progression and metastasis. The aim of this study is to screen out ideal markers for predicting progression and metastasis of early ESCC.Methods:The first part:surgically resected specimens from 24 patients with early ESCC(T1N0 stage), were studied. According to the survival time, they were divided into two groups. Patients in death group were 11 cases who died in 40 months and patients in survival group were 13 cases with at least 80 months duration. The genomic DNA isolated from tumor tissues were hybridized to Agilent CGH microarray. The second part:16 of 24 were analyzed for expression of microRNAs by TaqMan Human MicroRNA Arrays.16 cases were divided into two groups based on the same standard.Results:1. Many chromosome alterations were identified in our study, including gains of 1q,3q,5p,6p,7q/p,8q/p,9q, 11q,12p,14q,16p,17q/p,18p, 19q/p,20q/p,22q and deletions of 3p,4q/p,5q,6p,9p,11q,13q,14q,18q,19q,21q. Compared with death group, some copy number changes occurred more frequent in survival group. They were gains of 1q,3q,5p,7q/p,8q/p,9q, 11q,12p,14q,16p, 17q,19q,20q,22q and loss of 4q/p,5q,13q. Some small scale changes occurred in the two groups were significantly different. They included gains of lq21.3-24, 3q21.1-24,3q26.1-26.33,5p13.1-15.31,7p22,7q11.23,8p11.2,8q22.1-24.3, 11q12.1-13.5,12p12.1-13.33,14q23.1,16p12.1-13.3,17q25.1,19q13.12, 20q11.21-13.33,22q12.1-13.2 and losses of 4p,4q31-35,5q22.3 and 13q12.21-12.2. Moreover,109 candidate genes for distinguishing the two groups were identified.2. Many microRNAs were found to be differently expressed between two groups in our study, and four microRNAs were identified for distinguishing these two groups. They include miR-574-3p, miR-19b, miR-31 and miR-28-3p.Conclusions:There are some specific changes of chromosome copy number, genes and microRNAs which might contribute to the potency for progression and metastasis of early ESCC.In our study, we screened out some specific markers of molecular genetics to predict the progression and metastasis of early ESCC. These markers included 109 genes with copy number changes and four microRNAs. The results of our study find the scientific proof for correct estimation on therapy of early ESCC.更多还原