【作者】 牛坚；

【导师】 钱海鑫；

【作者基本信息】 苏州大学 ， 普通外科学， 2006， 博士

【摘要】 原发性肝癌是常见的消化系统恶性肿瘤,约占消化系统肿瘤的20%以上,其中肝细胞肝癌约占90%,其生物学特征为易复发、转移、预后差、死亡率高。尽管目前原发性肝癌的治疗方案(手术切除、介入治疗、生物治疗、激光及微波治疗等)日趋完善,但仍难以取得令人满意的疗效,患者生存期极短。因此研究肝癌的发病机理以及探讨新的可行的治疗方案具有极其重要的临庆意义。随着肿瘤学领域中基础研究的不断深入,人们发现在肿瘤组织中存在一些生长因子及其受体过表达的现象,为探讨肿瘤的发生机制奠定了基础。胰岛素样生长因子Ⅰ受体蛋白(IGFIR)是一种重要的促生长因子,它参与肿瘤细胞的生长、分化、转移、增殖等多方面的过程。因此IGFIR可能与肝癌的发生、发展有重要的相关性联系。目前,国内外针对IGFIR基因表达与肝细胞癌分化程度、生长方式、临床特点及转移活性的关系尚未进行深入系统的研究。针对肝癌的胰岛素样生长因子Ⅰ受体蛋白(IGFIR)利用小干扰RNA基因治疗尚无报道。本课题从三个角度进行了系统研究:①肝癌组织IGFIR基因表达与肝癌恶性程度、临床特点、转移活性的相关性和在肝癌细胞中的表达。②采用脂质体体外转染针对IGFIR的小干扰RNA,并建立稳定株观察转染后肿瘤细胞在体外的生长情况及产生的可能机制。③IGFIR小干扰RNA转染后肿瘤细胞体内生长情况观察。结果将为探讨以IGFIR为优选靶并结合小干扰RNA进行基因治疗,阻止肿瘤的发生和发展,提高生存率,延长生存期奠定理论基础。第一部分IGFIR在肝癌组织、肝癌细胞系中的表达及意义本研究对24对肝细胞癌(HCC),4例正常肝组织、4例胎肝组织以及14株肝癌细胞株用RT-PCR反应、免疫组化法、Western-blot分别检测IGFIRmRNA表达和蛋白表达的情况;分析肝癌IGFIR基因表达与肿瘤分期、转移等临床指标之间的关系。1通过RT-PCR方法检测24对人肝细胞癌、4例正常肝脏组织和4例胎肝组织IGFIRmRNA表达水平:发现肝癌组织IGFIRmRNA的表达明显高于癌旁组织,其中有20对下调,所占比率高达63%(P<0.01)。其中有7对接近有和无的关系。正常肝和胎肝中没表达。此外发现:瘤栓组更多还原

【Abstract】 Liver cancer is one of the common types of gastrointestinal tumors with poor prognosis metastasis and high mortality in which hepatocellular carcinoma(HCC)accounts for about 95%. Although the different available multi-modalities of therapies,including radical surgical operation、intervention therapy、laser therapy et al,have currently been used in the treatment for liver cancer,the prognosis of liver cancer has not been markedly improved. Therefore, it has an important clinic significance to study on the mechanism of the development and progression of liver cancer and the new strategies for the treatment of liver cancer with the molecular biology. With the development of the basic research on oncological fields, it was found that many tumors have the over expression in growth factors& their receptors, and progression of tumors depends on angiogenesis. The insulin-like growth factor (IGFIR)is an important factor which elicits the angiogensis in tumors. It takes an important role in the growth、differentiation、metastasis、proliferation of tumors. so IGFIR is considered to have key roles in the development of HCC. By now,the systemic researches on the relationship between the expression of IGFIR and histopathologic type, metastasis activity、angiogenesis of HCC have not been reported. And there are also no reports on gene therapy against angiogeneis of liver cancer. This study contains three parts of work: The first, the relationships between the expression of IGFIR in HCC and the tumor grade,tumor metastasis,angiogenesis of liver cancer. The second the inhibitory effect of IGFIR small interfere RNA on the growth of 7721 hepatocellular cell line (7721 cancer hepatocytes)in vivo. The third,the inhibitory effect of IGFIR small interfere RNA on 7721 cell in nude mouse.Part I: Study on the expression of IGFIR and its relationship to metastasis activity and angiogenesis of human HCC1 Expression of IGFIR gene in human HCC and normal liver tissue. The gene expression of IGFIR in 24 pair human HCCs and 4 normal liver were studied by RT-PCR method. The expressive rates of HCCs and normal tissue were 63% and 0% respectively. 7 paired samples are yes or no. Moreover, The levels of IGFIR mRNA (IGFIR mRNA/?-actin mRNA) were 105.9±6.8 in embolus, 120.3±6.3 in metastasis respectively. The difference between them are significant. The expression levels of IGFIR mRNA were positively correlated to pathologic grades、embolus and更多还原