抑制消减杂交法筛选原发性肝细胞癌中差异表达的基因及其生物学意义

为了筛选原发性肝细胞癌 (hepatocellularcarcinoma ,HCC)中差异表达的基因 ,以了解HCC发生发展的分子基础 ,选取了一例早期高分化肝癌标本作为材料 ,采用抑制消减杂交 (suppressionsubtractivehybridization ,SSH)技术 ,进行了前向及反向消减杂交 ,结合反向Northern印迹筛选 ,得到多个差异表达的基因 .对有意义的基因用半定量RT PCR检测了肝癌中的表达 .结果显示 ,PON2、hSRP1alpha、H4 1在大部分肝癌中表达升高 ,IGFBP1、ITIH1在早期癌症中 ,大部分癌的表达升高 ,在晚期癌症中则表达下降 .EGR1在大部分肝癌中表达降低 .研究表明 ,不同分化程度、不同临床分期的肝癌 ,有共同的或不同的基因表达发生改变 ,明确这些差异表达的基因谱 ,对于肝癌发生发展机理的阐明及肝癌的预防、诊断、治疗都有重要意义 .

Screening and Analysis of Differentially Expressed Genes in Human Hepatocellular Carcinoma Using Suppression Subtractive Hybridization

To understand the molecular basis of tumorigenesis and development of human hepatocellular carcinoma (HCC), suppression subtractive hybridization (SSH) in two directions were performed to identify genes that were differentially expressed in HCC,using an HCC sample of early stage with well differentiation status. Reverse Northern blot was performed to screen the truly differentially expressed genes,which resulted in identification of many genes. RT-PCR was used to detect the expression of some of the important differentially expressed genes in HCC samples. The results showed that the expression of PON2,hSRP1alpha and H41 were up-regulated in the majority of cancer tissues. Whereas the expression of IGFBP1 and ITIH1 were up-regulated in the early stage but down-regulated in the advanced stage. The expression of EGR1 was down-regulated in the majority of the cancer tissues. These findings suggested that there are common or different genes expression abnormalities in the HCC samples of different differentiation status and clinical stage. The identification of the gene expression profile in HCC will help us to clarify the mechanism of tumorigenesis and development of HCC and develop new targets for prevention, diagnosis and therapy.