应用抑制性消减杂交方法分析结肠癌相关基因

Analysis of Colon Cancer Related Genes by Suppression Subtractive Hybridization

目的：应用抑制性消减杂交（suppression substractive hybridization,SSH）方法分析、研究结肠癌相关基因。方法：在联合使用全反式维甲酸（all-trans-retinoic acid A,TRA）和1,25－二羟维生素D3[1,25-(OH)2D3]诱导大肠癌细胞株LoVo细胞分化的基础上，以大肠癌细胞提取的mRNA为模板合成的cDNA作为测试子，逆转细胞提取的mRNA为模板合成的cDNA作为驱赶子，进行SSH实验，研究LoVo细胞诱导前、后基因表达的差异。结果：首次证明ews基因和htert/hest2基因在大肠癌细胞中有表达，首次报道2个表达序列标签(expressed sequence tags,EST)（基因库登录号分别为AW266488和AW266490）。结论：大肠癌的发生是多基因参与的复杂过程。新EST提示诱导分化由多个基因参与，本文结果为阐明大肠癌的发病机制和诱导分化的机制以及筛选大肠癌候选基因提供了实验依据。

Objective：This study was designed to investigate the colon cancer related genes by suppression subtractive hybridization(SSH). Methods：Combinative all-trans-retinoic acid (ATRA) and 1,25-dihydroxy vitamin D3 [1,25-(OH)2 VD3] were used to induce the normal differentiation of colon cancer LoVo cells, and on this basis, with cDNA from control cancer cells as tester and cDNA from induced cancer cells as driver, SSH was used to investigate the difference of expressed genes of LoVo cells before and after induction. Results：ews and htert/hest 2 genes were first found expressed in colon cancer, 2 new expressed sequence tags(EST) were first found (their GenBank accession numbers are AW266488 and AW26490). Conclusion：The pathogenic mechanism of colon cancer is a complex process with multiple genes involved. New ESTs suggest that many genes take part in inducing differentiation. The results provide the bases for partially explaining the pathogenic mechanism of colon cancer, the mechanism of inducing differentiation and screening the colon cancer candidate genes.