用SSH法筛选喉肿瘤组织与其周围正常组织的差异表达基因

Suppression subtractive hybridization to identify differentially expressed gene between laryngeal tumor and the normal tissue adjacant to the tumor

目的 :筛选喉肿瘤组织与其周围正常组织的差异表达基因 ,建立消减cDNA文库。方法 :用抑制性消减杂交 (SSH)法建立消减cDNA文库 ,随机挑选 9个克隆通过进行反向Northern杂交证实 ,测序所得序列在GeneBank中作blastn分析发现。结果 :除 1个克隆为接头相连外 ,其余 8个克隆代表了 6个基因和 1个免疫球蛋白重链区。 6个基因除 1个代表功能不明的新抗体外 ,其他基因分别为 :肿瘤坏死因子、角质素、肌动蛋白、肌球蛋白以及碱性磷酸酶。结论 :SSH法筛选差异表达基因真实可靠。运用该法筛选的基因在肿瘤的发生发展、肿瘤的转移、肿瘤免疫和肿瘤标志中发挥重要作用。

Objective:To screen the differently expressed genes between laryngeal tumor and the normal tissue adjacant to the tumor and construct the subtractive cDNA library.Method:The subtractive cDNA libra- ry was contructed with the suppression subtractive hybridization(SSH)technique.Nine random clones are made as probes for the reverse Northern hybridization,after sequenced.The sequences are compared with known sequences using blastn against GenBank across the Internet.Result:Except one clone ligated by two adaptor,the others represented six gene and one IgG heavy chain constant region.Among the six genes,one was a new kind of antibody whose function was not understood,the others stood for genes as follows:keratin,β-actin,myosin,tumor necrosis factor,alkaline phosphatase.Conclusion:Screening the differently expressed gene with SSH is reliable.These genes play a pivotal role in tumorigenesis,tumor promotion,tumor metastasis,tumor immunity and tumor marker.