肝癌形成过程中基因表达谱及转录核因子-κB动态改变

Dynamic alteration of gene expression profiling and nuclear factor-κB during development of hepatocellular carcinoma

目的观察肝癌(HCC)形成过程中基因表达谱及转录核因子-κB(NF-κB)的动态改变。方法雄性SD大鼠以2-乙酰氨基芴(2-FAA)喂饲制备肝癌模型,以全基因组芯片分析基因表达谱的动态变化,经病理组织学(HE染色)分析肝细胞的形态学变化,在癌变不同时期定量观察NF-κB动态变化,并分析了人肝癌及非癌组织中总RNA及NF-κB表达水平。结果鼠肝癌形成过程中,经含有28 000个基因芯片分析发现,早期2 898个基因上调,晚期3 208个基因上调。正常对照肝NF-κB表达水平较低,在诱癌后肝细胞发生颗粒样变性、不典型增生、高分化肝细胞癌形成过程中,随肝癌进展NF-κB表达呈梯度增加,在肝癌发展不同阶段NF到-κB表达水平均显著高于对照组,并与肿瘤坏死因子-α表达高度正相关。人肝癌组NF-κB表达,也明显高于自身对照的非癌组织(P<0.01)。结论多种基因参与肝细胞的癌变过程,其中NF-κB信号转导途径激活起起重要作用。

Objective To investigate the gene expression profiling and kinetic expression of hepatic nuclear factor- Kappa B （NF-κB） during the course of hepatocyte carcinogenesis. Methods Rat hepatoma models were made with N- （2-fluorenyl） acetamide （2-FAA） on male Sprague-Dawly （SD） rats. Gene expression profiling was analyzed by rat DNA array and morphological changes were observed by pathological examinations （ HE staining） in rat model. Dynamic Alterations of NF-κBB at different stages of rat HCC model, and expressions of NF-κB in human HCC and their non-HCC tissues were quantitatively analyzed by ELISA, respectively. Result The up-regulated expressions of 2 898 genes at early stages and 3 208 genes at end stages were found by DNA array analysis among total 28 000 genes during the course of HCC formation and development. The hepatocytes showed vacuole-like denaturation at the early stages, then hyperplastic nodal appearance at middle stage, and finally progression to tubercles of cancerous nest with highly differentiated hepatocyte cancers during development of rat hepatoma. An increasing tendency of liver NF-κB expression was found from normal liver to precancerous to cancerous tissues during rat hepatoma development and comparison with lower expression in normal controls, and closely relationship was found between NF-κB and TNF-α. The level of NF-κB in tissues of human HCC was higher than that in their paracancerous tissues （P 〈 0.01 ）. Conclusion Muhigenes would participate in the course of hepatocyte canceration and the reactivation of NF-κB signal transduction pathway play an important role in HCC.