摘要
目的:运用微阵列芯片技术研究喉鳞状细胞癌及癌旁正常黏膜中miRNA表达谱差异,探讨miRNA与喉癌发病的关系。方法:提取8例喉癌患者的癌组织及其对应癌旁正常黏膜中的miRNA,分别与哺乳动物miRNA芯片V3.0杂交,扫描荧光信号,对癌组织、正常黏膜进行差异基因筛选和聚类分析;采用实时定量RT-PCR法验证部分差异表达的miRNA。结果:共筛选出47个喉癌相关的差异表达miRNAs基因:24个在喉癌组织中表达下调,23个表达上调。其中下调5倍以上的有miR-1,miR-486-5p,miR-206,miR-487a,miR-375,miR-422a,miR-144,miR-384,miR-378,miR-133a;表达上调3倍以上的是miR-93,miR-31和miR-20b。在喉癌差异表达的miRNA中有13个成5簇共表达,分别位于8,13,14,18和X染色体上。荧光定量RT-PCR结果与miRNAs芯片的结果较符合。结论:一些miRNA可能参与喉癌的发生的调控。
Objective：To detect the expression of microRNAs （ miRNAs） in laryngeal carcinoma and adjacent normal tissue using microarray,and to discuss the relationship between miRNAs and laryngeal carcinoma. Method：miRNA were extracted from 8 cases of laryngeal cancer tissue and its adjacent normal tissue. miRNA identification were performed by microarray of miRNA hybridization and cluster analysis was used with Significance Analysis of Microarrays （ SAM,version 2. 1） and Cluster3. 0 software. miRNA were confirmed by real time quantification RT-PCR with RNA-tailing and primer extension. Result：Totally 47 different miRNAs were found expressed in laryngeal cancer,with 23 of miR-NA expression were up-regulated and 24 of miRNA expression were down-regulated. The expression of miR-1,miR-486-5p,miR-206,miR-487a,miR-375,miR-422a,miR-144,miR-384,miR-378,miR-133a were down-regulated by 5 multiple and while expression of miR-93,miR-31,miR-20b were up-regulated by 3 multiple. There are 5 miRNA clusters with coexpression in laryngeal cancer tissue and located on chromosome 8,13,14,18 and X. Moreover,RT-PCR analysis demonstrated that there was no significantly difference of miRNA expression between microarray and RT-PCR. Conclusion：The different expression of miRNA may play an important role in the pathogenesis and development of laryngeal cancer.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2010年第12期535-538,共4页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
基金
吉林省科技发展计划项目(No:200905197)
吉林省卫生厅重点资助项目(No:20082005)
关键词
喉肿瘤
微小RNA
基因表达
laryngeal neoplasms
microRNA
gene expression
