应用抑制性消减杂交技术克隆人肺鳞癌差异性表达基因

目的 克隆人肺鳞癌肿瘤差异性表达基因。方法 应用抑制性消减杂交技术（suppression subtractive hybridization,SSH)构建人肺鳞癌cDNA消减杂交文库，筛选后挑选阳性克隆进行测序，并在Gen Bank数据库中进行同源性比较，最后经RT－PCR验证。结果 成功克隆10个差异基因片段，其中2个为新基因（GenBank登录号分别为：C20：AF363068；C34：AY032661）。结论 抑制性消减杂交技术是克隆差异表达基因及发现新基因的有效方法。

Molecular cloning of genes differentially expressed in human lung squamous cell carcinoma using suppression subtractive hybridization

Objective To clone and identify genes differentially expressed in human lung squamous cell carcinoma (LSCC). Methods A subtracted cDNA library of human LSCC was constructed by suppression subtracted hybridization (SSH) method. After screening, the subtracted library clones representing mRNAs that were truly differentially expressed in LSCC but not in its adjacent non cancerous tissues were selected to identify by RT PCR and DNA sequencing were performed. Nucleic acid homology searches were performed using the BLAST program. Results By this technique, 10 differentially expressed gene cDNA fragments of LSSC were obtained. Two were novel and eight were already known genes. Conclusion SSH is a useful technique with high sensitivity for the detection of differential genes expression in LSCC and an effective method to clone novel genes.