翻译起始因子eif3b在肝癌发生中作用机制分析-mechanism analysis of translation initiation factor eif3b in hepatocarcinogenesis.docxVIP

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翻译起始因子eif3b在肝癌发生中作用机制分析-mechanism analysis of translation initiation factor eif3b in hepatocarcinogenesis.docx

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翻译起始因子eif3b在肝癌发生中作用机制分析-mechanism analysis of translation initiation factor eif3b in hepatocarcinogenesis

中文摘要中文摘要 IV IV 第三部分：EIF3B 通过影响 MAPK 信号通路中关键基因发挥促癌作用目的：初步探讨 EIF3B 在肝细胞癌发生过程中的作用机制方法：采用全基因表达谱芯片筛选 EIF3B 敲减后下游应答基因，通过生物信息学分析，对应答基因进行 GO，Pathway 分析，挑选 MAPK 信号通路中关键基因进行 Q-PCR 和 Western blotting 验证。结果：以 FC 2.0 或 FC 0.5，P0.05 为筛选标准，表达谱基因芯片结果显示 EIF3B 敲减后 SMMC-7721 细胞基因发生改变的有 390 个，其中上调的有 196 个，下调的有 194 个； KEGG 与 BIOCARTA 通路富集分析， MAPK 信号通路中有 10 个关键基因的表达水平发生改变；Q-PCR 和 Western blotting 结果显示 MAPK 通路中 Dusp5，EGFR，ZAK， PPM1A 的表达水平发生改变。结论：EIF3B 通过影响 MAPK 信号通路中关键基因在肝细胞癌发生过程发挥促癌作用。关键词：EIF3B；肝细胞癌；发病机制；丝裂原活化蛋白激酶信号转导通路。 Abstract Abstract PAGE PAGE VI ABSTRACT Part One EIF3B is overexpressed in hepatocellular carcinoma. Objective: To investigate the expression of EIF3B in hepatocellular carcinoma tissues and hepatocellular carcinoma cell lines and to explore the association between EIF3B expressions and clinicopathological features of hepatocellular carcinoma. Methods：Expressions of EIF3B mRNA in hepatocellular carcinoma tissues and hepatocellular carcinoma cell lines were determined by real-time PCR technology, EIF3B protein of paraffin-embedded specimens were examined by immunohist- ochemistry. Results：Results of real-time PCR showed that EIF3B mRNA level was higher in hepatocellular carcinoma tissues and cell lines. A significant overexpression of EIF33B was found in hepatocellular carcinoma lesions compared to the matching adjacent non-tumor tissues. High expression of EIF3B was significantly correlated with advanced tumor-node-metastasis stage and poor survival rates, but had no relationship with age, gender, tumor size, and tumor differentiation. Conclusions：Both EIF3B mRNA and protein expressions are higher in HCC tissues. EIF3B may play a role in hepatocarcinogenesis. Keywords: hepatocellular carcinoma; EIF3B; real-time PCR; Immunohi- stochemistry Part Two EIF3B silencing changes the phenotype of hepatocellular carcinoma. Objective: To investigate the effect of EIF3B gene silencing by lentiviral vector mediated siRNA and its effect on biological behavior in hepatoce