人胃印戒细胞癌组织差异表达基因克隆的研究

目的:分离胃印戒细胞癌组织差异表达基因,探讨人胃印戒细胞癌发生的分子机制。方法:采用抑制性消减杂交技术(SSH),构建人胃印戒细胞癌组织cDNA 消减文库;随机挑选部分阳性克隆测序,与GenBank中的已知序列进行基因同源性分析,并进一步探讨基因功能。结果:成功构建高消减效率的人胃印戒细胞癌组织cDNA消减文库,并分离出多个差异表达基因片段:其中1 条与染色体序列相似,可能代表未知的新基因序列,已被GenBank 接收(注册号:CN446913);其他多为已知的肿瘤相关基因的部分片段。结论: 1 )SSH技术是分离差异表达基因的有效方法;2)胃印戒细胞癌的发生发展与多种基因表达异常有关;

Study on cloning of differentially expressed genes in stomach signet ring cell carcinoma

OBJECTIVE:To isolate genes expressed exclusively or preferentially in human stomach signet ring cell carcinoma in an effort to explain the mechanisms in the development of the tumor. METHODS:Suppression subtractive hybridization (SSH) was performed to set up a subtraction cDNA library of specially or highly expressed genes in gastric signet ring cell carcinoma. Some clones were randomly picked and sequenced. These genes were compared with known sequences in the public databases of GenBank/EMBL/DDBJ by using NCBI BLAST. The functions of these genes on the development of tumor were analysed.RESULTS:1)A subtracted cDNA library of differentially expressed genes in human gastric signet ring cell carcinoma was constructed successfully with SSH and T/A cloning techniques. 2)Some differentially expressed gene fragments were cloned: one was similar to chromosome sequence and submitted to ESTdatabase (dbEST) at NCBI as an EST of a new given gene (Accn:CN446913),the others were from previously described genes,a large proportion of which had been reported to be linked with tumor.CONCLUSIONS:1)SSH is an effective way to isolate differentially expressed genes. 2)Genes expressed differentially may play an important role in the development and growth of human gastric signet ring cell carcinoma.