用荧光差异显示法分离新的肝细胞癌相关基因

Identification of new hepatocellular carcinoma related genes by fluorescent differential display

目的 :寻找新的肝细胞癌 (hepatocellularcarcinoma ,HCC)相关基因。方法 :应用荧光 差异显示 (fluorescentdifferentialdisplay)技术分析人HCC、配对非癌肝、胎肝、正常肝等组织之间的基因差异表达 ,差异表达cDNA片段 ,经测序后 ,再用RNA狭缝杂交对部分差异片段在上述 4种组织中的表达进行验证。结果 :共得到差异表达cDNA片段 110条。经狭缝杂交筛选 ,获得了 4条阳性片段 ,未知cDNA片段L2和与人醛缩酶B 99.5 %同源的cDNA片段L7在部分HCC中特异性低表达 ,未知cDNA片段LC2 7和与人Nip3基因 99.4%同源的cDNA片段LC2 0在部分HCC中特异性高表达。结论 :确认了 4条HCC相关基因。其中L2和人醛缩酶B基因可能为阻抑HCC发生、发展的基因 ,而人Nip3基因和LC2 7可能为促进HCC发生、发展的基因。L2和LC2 7为目前未知的新基因。

Objective: To identify new hepatocellular carcinoma (HCC) related genes. Methods: Fluorescent differential display (Fluoro DD) was used to examine difference in the mRNA compositions of human HCC, paired nontumor liver, fetal liver and normal liver tissues. Some of differentially expressed cDNAs were sequenced and their expression patterns were further detected by RNA slot blot. Results: 110 differentially expressed cDNA fragments were obtained. The results from RNA slot blot showed that the transcription of L2 and L7 (99.5% identity with human aldolase B gene) was down regulated in some cases of HCC, while the transcription of LC20 (99.4% identity with human Nip3 gene) and LC27 was up regulated in some cases of HCC. Conclusion: Four HCC related genes have been identified. L2 and human aldolase B gene may act as the candidates for HCC suppression related genes, LC27 and human Nip3 gene may act as the candidates for HCC promotion related genes, respectively. L2 and LC27 are new genes so far.