应用抑制性消减杂交技术克隆人肺鳞癌差异性表达基因  

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

作　　者：沈晨阳[1] 刘军[1] 王丹蕾[1] 赵辉[1] 姜冠潮[1] 王俊[1] 张国良[1] 

机构地区：[1]大学人民医院胸外科,北京100044

出　　处：《中国肺癌杂志》2002年第1期10-13,共4页Chinese Journal of Lung Cancer

基　　金：国家自然科学基金 ( 39970 72 9)资助

摘　　要：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 (suppressionsub tractivehybridization,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GenBank数据库中进行同源性比较 ,最后经RT PCR验证。结果　成功克隆 1 0个差异基因片段 ,其中 2个为新基因 (Gen Bank登录号分别为 :C2 0 :AF3630 68;C34 :AY0 32 661 )。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.

关 键 词：差异性表达基因 抑制性消减杂交技术 肺癌状细胞癌 基因克隆 肺肿瘤 互补DNA 

分 类 号：R734.2[医药卫生—肿瘤]