生物芯片和rna干扰技术在肝癌和法医学研究中的应用遗传学专业论文.docxVIP

复旦大学博士学位论文 复旦大学博士学位论文 生物芯片和RNA干扰技术在肝癌和法医学研究中的应用 我们系统的结合基因芯片、组织芯片及RNA干扰分析来鉴别和验证了与肝癌 的发生发展相关的一些新的生物学标记物。我们鉴别出了一个能够预测肝癌样本 分化程度及术后生存情况的9基因判别式。我们的研究同时表明，MCM2和CCNBl 是与肝癌的发生发展相关的潜在的生物学标记物，它们有希望成为肝癌治疗的分 子靶点。我们的研究有助于进一步阐明肝癌的分子机制，为鉴别新的治疗靶点提 供线索。 此外，我们开发出了适用于我国人群的SNP芯片，能够检测45个SNP位点， 个体识别率达到10。9，灵敏度与当前STR的基因型检测技术相当，成果可望应用 于各个法医物证学鉴定机构(亲子鉴定和个体识别)，使法医DNA分析技术进入 一个新的领域。 关键词：肝癌·生物学标记物·cDNA芯片·基因表达谱·组织芯片·RNA 干扰·MCM2·CCNBl·SNP检测芯片 AbstractHuman Abstract Human hepatocel lular carcinoma(HCC)i S one of the most common cancers worldwide．Currently，there is no effective therapy for most HCC patients． It is critical to identify and validate novel anticancer targets in HCC． It is generally accepted that eDNA microarrays，tissue microarray(TMA) and RNA interference(RNAi)are useful tools for identification and val idation of biomarkers in disease research． The purpose of this study was to identify and val idate novel diagnostic and therapeutic targets in HCC． We analyzed gene expression profi 1es not only between 33 HCCs and their corresponding noncancerous liver tissues，but also between 25 HCCs and pooled normal 1iver tissues using cDNA microarrays containing 12800 genes．Functional analysis of differentially expressed genes involved in HCC carcinogenesis and tumor progression revealed that up—。regulated genes are mainly associated with cell cycle，while down—regulated genes are mainly associated with immune response．We detected two regions of cytogenetic changes only in poorly——differentiated HCCs using the expression data．We identified two novel human genes and other twenty genes which have not been reported in HCC before．They were significantly up-regulated in at least 50％HCCs and might be potential biomarkers for HCC diagnosis．We identified a 9-gene expression signature，which was able to predict differentiation degree of HCC samples and was a more powerful predictor of the survival of HCC patients than standard systems based on clinical and histologic criteria．Among the 9 most discriminatory