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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 differentially expressed genes in laryngeal neoplasms and their surrounding normal tissues and establish subtractive cDNA libraries. Methods: Subtracted cDNA library was established by SSH. Nine clones were randomly selected and confirmed by reverse Northern blotting. The sequences were sequenced and analyzed by blastn in GeneBank. Results: All but one of the clones represented six genes and one immunoglobulin heavy chain region, except one clone was ligated. 6 genes in addition to a representative of unknown function of new antibodies, the other genes were: tumor necrosis factor, keratin, actin, myosin and alkaline phosphatase. Conclusion: SSH method of screening differentially expressed genes is reliable. The gene screened by this method plays an important role in tumorigenesis, tumor metastasis, tumor immunity and tumor marker.