食管癌中高表达的钙结合蛋白1基因的序列分析及意义

目的:筛选食管癌中差异表达的基因,为进一步了解食管癌发生的分子机制奠定基础.方法: 利用荧光差异显示技术(DD-PCR)获得差异片段,对这些片段进行克隆和序列分析.通过在GenBank中同源性检索,查找差异片段相对应的同源基因.利用定量PCR检测验证该基因表达的差异性,并对该基因的结构进行预测和分析.结果:通过DD-PCR得到差异片段中的一个片段对应于人类钙结合蛋白1基因(Cabin1).荧光定量PCR技术检测结果表明钙结合蛋白1基因在食管癌组织中的表达量高于其对应的癌旁组织,在食管癌细胞株TE1中的表达量高于其他被测试的5种肿瘤细胞株.生物软件分析表明:该基因的读码框长6 663bp,编码2 220个氨基酸,至少有4种剪切方式.结论:Cabin1在食管癌组织及食管癌细胞中异常表达,可能在食管癌发生过程中起着重要的作用.

Sequence analysis and its significance of calcium binding protein 1 gene expressed highly in esophageal cancer

Objective: To further investigate the molecular mechanism of esophageal carcinogenesis and screen out the genes expressed differently in esophageal cancer.Methods: mRNA differential display(DD-PCR) was employed to search differently expressed fragments,some of which were cloned and sequenced.By homology analysis in GenBank,corresponding homologous genes of those fragments were found.The corresponding genes of those differently expressed fragments were validated by Real-time quantitative PCR. Results: By DD-PCR,one EST was identified to be calcium binding protein 1(Cabin1).The result of real-time quantitative PCR showed that the expression level of Cabin1 in esophageal cancer tissues was significantly higher than that in normal tissues,and its expression level in esophageal cancer cell line was also higher than that in five other cancer cells from lung cancer and Leukaemia.The ORF of Cabin1 was 6663 base pair long and encodes 2220 amino acids,and the gene had at least four splicing forms. Conclusion: The over-expression of Cabin1 in esophageal cancer may indicate an important role in esophageal carcinogenesis.