喉鳞癌及癌旁正常组织miRNA差异表达分析  

Analysis of differencial miRNA expression in squammous cell carcinoma of the larynx and adjacent normal tissues

作　　者：魏明辉[1,2] 张宗敏[3] 李琳[4] 高燕宁[4] 唐平章[3] 

机构地区：[1]深圳市第二人民医院 [2]深圳大学第一附院耳鼻喉科,广州深圳518035 [3]中国医学科学院肿瘤医院头颈外科,北京100021 [4]中国医学科学院肿瘤研究所病因与癌变研究室,北京100021

出　　处：《中华肿瘤防治杂志》2014年第7期506-508,513,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的：应用基因芯片技术，研究喉鳞状细胞癌中miRNA的表达谱，筛选喉鳞癌和癌旁正常中组织差异表达的miRNA，寻找可能的分子生物学指标。方法：选取2008—06—01—2009-04-30中国医学科学院肿瘤医院8例喉鳞癌及其配对的癌旁正常组织（距肿瘤组织〉1cm），Trizol法提取总RNA，经Cy3荧光标记后与含有人723条miRNA探针的芯片进行杂交，筛选在喉鳞癌和癌旁正常组织中差异表达的miRNA。结果：喉鳞癌及癌旁组织中提取的总RNA均符合芯片杂交要求，RNA完整性评分1．8～2．1；共有132个miRNA在8例喉鳞癌患者中均有表达。喉鳞癌和癌旁正常组织中差异表达的miRNA有53个，其中22个出现上调，31个出现下调，差异表达≥5倍的miRNA共有8个，上调的有miR-196b、miR-196a、miR-7和miR-183，下调的有miR-375、miR-139—5P、miR-338-3p和miR-370，其中miR-196b、miR-196a和miR-375差异表达≥8倍。采用TargetScan、Pictar、miRanda和MirTarget2等4种算法数据库预测这8种差异表达的miRNA的靶基因，4种数据库中均预测到的靶基因共183个。结论：通过基因芯片的方法，筛选到喉鳞癌和癌旁组织中差异表达的miRNA，对其中明显差异表达的miRNA及其靶基因的深入研究有可能找到新的喉鳞癌分子肿瘤学标志，为进一步研究喉鳞癌发病机制打下基础。OBJECTIVE：To study the expression profiles of miRNA in squamous cell carcinoma of the larynx by microar-ray, analyze the altered miRNA expression of suqamous cell carcinoma of the larynx compared with adjacent normal tissue and explore the potential molecular signature specific for the laryngeal carcinoma. METHODS：Total RNA was extracted in 8 squamous cell carcinomas of the larynx and matched adjacent normal tissue by Trizol method,labeled with Cyanine3-pCp and hybridized with human 723 miRNA microarray. Differential miRNA expression between laryngeal carcinomas and ad-jacent normal tissues was determined. RESULTS：The total RNAs extracted from tissues of squamous carcinoma and adja- cent normal tissues were qualified for gene microarray hybridization. Their RNA Integrity Number ranged from 1.8 to 2.1. Totally 132 miRNAs were detected in all the 8 samples of laryngeal carcinorma. Fifty-three miRNAs were determined to be differentially regulated in the squamous cell carcinoma when compared with the normal adjacent tissues. There were twenty-two miRNAs upregulated and thirty-one downregulated. Eight miRNAs were found to be more than five fold alter-ation,four of them were upregulated including miR-196b,miR-196a,miR-7 and miR-183. The other four miRNAs inclu-ding miR-375,miR-139-5p, miR-338-3p and miR-370 were downregulated. Three of them including miR-196b, miR-196a and miR 375 were more than eight fold alteration. The Target genes of the 8 differentially expressed miRNAs were predic- ted by four softwares of TargetScan, Pictar,miRanda and MirTarget2. There were 183 target genes which were predicted in all the four softwares. CONCLUSIONS.. The differentially expressed miRNAs belween squamous cell carcinoma of the larynx and adjacent normal tissues are screened by microarray. The advanced research to the differentially expressed miR NAs and their targel genes may help us find new molecular marks for the squamous cell carcinomas of the larynx and, which may prepare a foundation for the further studies in the squ

关 键 词：喉肿瘤 肿瘤 鳞状细胞 基因芯片 分子生物学 

分 类 号：R739.65[医药卫生—肿瘤]