摘要
目的 分析人肺鳞癌组织和正常肺组织中差异表达的肿瘤相关基因的表达状况 ,探讨其在肺癌发生、发展中的作用。 方法 对经抑制性消减杂交技术 (suppressionsubtractivehybridization ,SSH)获得的部分阳性克隆的差异表达基因 ,采用半定量RT PCR方法在 12例肺鳞癌标本中进行验证 ;对其中的部分新基因进行NorthernBlot检测。 结果 共获得 10个差异表达基因片段 ,其中 6个为已知基因片段 ,2个为结构已知但功能未知的基因片段 ,2个可能为新基因EST片段(GenBank登录号 :C2 0 /AF36 30 6 8;C34/AY0 32 6 6 1)。经检测 ,包括新基因AY0 32 6 6 1在内的 6个基因在肺鳞癌肿瘤组织中表达下调 ;包括新基因AF36 30 6 8在内的 4个基因在肺鳞癌肿瘤组织中表达上调。 结论 所分析的差异表达基因均与肿瘤的发生和机体的免疫机制有关。
Objective To identify genes differentially expressed in human lung squamous cell carcinoma (LSCC). Methods A subtracted cDNA library of human LSCC constructed by using suppression subtracted hybridization (SSH) method was screened. Clones representing mRNAs that were truly differentially expressed in LSCC but not in its adjacent non-cancerous tissues were identified by semi-quantitative RT-PCR in 12 patients with LSCC. Partial novel genes were detected by Northern blot. Results Ten differentially expressed gene cDNA fragments of LSSC were obtained by SSH. Among them six were known genes; two sequences were identified but their functions were unknown (hypothetical protein); two were novel (GenBank accession numbers AF363068 and AY032661, respectively). The results from semiquantitative RT-PCR showed that the transcription expression level of PPP1CB, caluminin, S100A2, HSNOV1, OCIA and AY032661 was down-regulated in some LSCC cases, while the transcription of HSP90, ferrintin, gp96 and AF363068 was up-regulated in others. Conclusions Six known genes identified by SSH technique have been implicated in the pathogenesis of lung carcinogenesis, or they are involved in immunological defense mechanism in human body. Two hypothetical proteins probably also play an important role in the pathogenesis of lung cancer. The function of the two novel genes in lung carcinogenesis are under investigation.
出处
《中华外科杂志》
CAS
CSCD
北大核心
2003年第5期368-371,共4页
Chinese Journal of Surgery
基金
国家自然科学基金资助项目 (3 9970 72 9)
关键词
肺鳞癌
肿瘤相关基因
基因表达
基因克隆
半定量RT-PCR
验证
Oncogenes
Oligonucleotide array sequence analysis
Carcinoma, non-small-cell lung
Reverse transcriptase polymerase chain reaction
