【作者】 郑江华；

【导师】 王广义；

【作者基本信息】 吉林大学 ， 外科学， 2010， 博士

【摘要】 原发性肝癌是世界范围内最常见的恶性肿瘤之一,我国是全球肝癌发病率最高的地区,肝癌术后高的复发率是影响患者预后的最主要因素。目前开展的针对肝癌生物学特性的研究,有可能为肝癌患者的临床治疗带来新的希望。基因芯片是近年来出现的一种高通量、快速、平行核酸序列测定及定量分析技术。基因芯片技术的出现,为在全基因组范围内寻找与肿瘤发生、发展相关的基因变化提供了一个强有力的手段,它只用一次实验、一次杂交反应就能在全基因组范围内同时分析待测样本中成千上万个基因的表达情况及其相互关系,这一巨大优势是其他分子生物学技术所无法比拟的。2003年1月开始,我们开始建立和维护一个信息化管理的符合基因组学研究的肝癌数据标本库,接受肝癌切除手术的患者在签署知情同意书后,进入肝癌数据标本库资料收集系统。至2009年12月,共有304例患者组织标本,126例血液样本,114例石蜡标本进入肝癌数据标本库资料收集系统。对标本定期随机抽样进行RNA提取鉴定符合相关分子生物学研究的要求。信息化管理的肝癌数据标本库为多个科研项目提供支持。我们从建立的肝癌标本库中各随机选取10例早期复发组和10例非早期复发组患者中进入本研究。我们利用Affymetrix基因芯片技术在全基因组范围内分析比较早期复发和非早期复发肝癌患者基因表达谱差异,进而通过数据挖掘筛选与肝癌早期复发相关基因。肝癌早期复发是一个多步骤、多因素参与的复杂过程,是多基因协同作用的结果。筛选到与肝癌早期复发密切相关的分子靶标,为肝癌的生物治疗提供新的靶点和思路,以此建立一组与肝癌早期复发密切相关的分子标签,进行更准确的分子学分类分型,并将为患者提供预后的判断和个体化的治疗。进一步利用免疫组化验证差异表达基因SOX4和CD24在早期复发组和非早期复发组肝癌之间的表达水平的差异,发现SOX4和CD24基因与肝癌的早期复发密切相关,SOX4和CD24基因高表达有可能促进了肝癌的侵袭和转移,并可能作为预防和治疗肝癌早期复发的分子靶标。更多还原

【Abstract】 PartⅠEstablishment and maintenance of HCC tissue and data bank by information managementObjective: To explore how to establish and maintenance HCC data bank of information management.Methods:HCC patients after operation were entered into HCC data bank, after signed informed consent. Fresh tissues were stored in liquid nitrogenSome tissue embedded in paraffin and the blood samples were collected. The collected clinical data and specimens is of information management.Resultes:304 cases of tissue specimens, 126 cases of blood samples, 114 cases of paraffin specimens of HCC were entered into HCC data bank. HCC data bank showed be suitable for molecular biologic research.Through information management, HCC data bank provide support for many research projects.Conclusion: Through establish and maintenance HCC data bank of information management, a good foundation is made for study of HCC.Part II Identification of differentially expressed genes associated with early recurrence of HCC using genechip technologyObjective: To identify the differential expression genes associated with early recurrence after curative resection for HCC using genechip technology.Methods: 20 cases selected from ER group and nER group randomly were entered into this study. The cRNAs were hybridized with Affymetrix genechip. The picture signals were analyzed with GCOS1.2. GeneSpring 10.0 analysis software screened differential gene. The differential expression genes identified by GO analysis, cluster analysis, principal component analysis and pathway analysis.Resultes: There is 2214 differential expression genes analyzed by the GeneSpring software. GO analysis found the differential expression genes were associated with 28 GO terms.The 20 cases could be divided into ER and nER correctly by using clustering analysis and principal component analysis. Pathway analysis showed that differential expression genes participated in apoptosis, transcription, proliferation, processes, and differential expression genes and may be involved in the process of tumor recurrence.Conclusion: The application of Affymetrix genechip technique is a powerful strategy to identify differential expression genes associated with early recurrence for HCC. The further study of the differential expression genes may found molecular mechanism of early recurrence of HCC and screen some new potential therapeutic targets for HCC.PartⅢExpression of SOX4 and CD24 and early recurrence of hepatocellular carcinoma: validation of their relationship and clinical significanceObjective: To explore study the relationship between expression of SOX4 and CD24 gene and early recurrence after curative resection of HCC, and its clinical significance.Methods: SOX4 and CD24 expression of HCC in 60 patients and normal liver was studied with the methods of immunohisto- chemisty. Resultes: The positive expression of SOX4 and CD24 in HCC was higher than in normal liver. The positive expression of SOX4 and CD24 in HCC of ER after curative resection was higher than HCC of nER. Conclusion: SOX4 and CD24 was confirmed be associated with early recurrence of HCC after curative resection. Up-regulation of SOX4 and CD24may contribute to early recurrence potential of HCC. SOX4 and CD24 may be developed as a new and effective molecular target for HCC in future. The detection of SOX4 in HCC might sever as an important parameter for estimating the prognosis.更多还原