食管鳞状细胞癌转移相关miRNAs差异表达谱分析  

The analyzation of miRNAs screening expression in esophageal squamous cell carcinoma

作　　者：周萍[1] 董贺[1] 刘群[1] 孙青[1] 

机构地区：[1]山东省千佛山医院,山东济南250014

出　　处：《济宁医学院学报》2012年第6期387-391,401,共6页Journal of Jining Medical University

基　　金：山东省自然科学基金面上项目(ZR2012HM085)

摘　　要：目的应用microRNA表达谱芯片技术,选取有详细临床病理数据及随访资料的食管鳞状细胞癌组织及相应正常食管粘膜组织,分析食管鳞癌患者有淋巴结转移组和无转移组差异性表达的miRNA。方法选取10例新鲜冻存的食管鳞癌组织及其相应的正常食管粘膜组织,使用microRNA芯片(miRCURY LNAmicroRNA芯片,Exiqon公司),筛选出食管鳞状细胞癌组织中差异表达的miRNA并进行聚类分析;运用实时定量逆转录聚合酶链反应(real time RT-PCR)验证结果的可靠性;最后通过生物信息学分析预测其靶基因。结果有淋巴结转移组与无淋巴结转移组相比较,差异表达的miRNA共有40个,其中表达上调者miRNA 22个,表达下调者miRNA 18个,经过聚类和PAM分析及real-time RT-PCR技术验证,筛选出二个具有组织特异性表达的miRNAs(miRNA-612和miRNA-583)。miRNA-612的表达上调及miRNA-583的表达下调在有淋巴结转移和无淋巴结转移食管癌中差异具有显著意义。结论通过miRNA芯片发现的相关差异性表达的miR-NA,有可能成为食管鳞状细胞癌淋巴结转移特征性的分子标志物,并为进一步认识和研究miRNA在食管鳞癌发生发展中的作用,寻找并鉴定与临床诊治相关的靶分子奠定基础。Objective We selected the esophageal squamous cell carcinoma cases with detailed clinical-pathological data and follow-up information.The miRNA expression profiles with miRNA microarray were investigated.The expression of miRNAs screening was analyzed between lymph node metastasis group and non-metastasis group.Methods 10 cases of fresh frozen samples of esophageal squamous cell carcinoma tissues and their normal tissues were selected.miRNA microarray（miRCURY LNA ？ microRNA chip,Exiqon company）was used to detect differential expression of miRNAs.And the expression of miRNAs screening was analyzed.Real time quantitative PCR was applied to verify the reliability of miRNA array results.The target genes were predicted through the database of bioinformatics analysis.Results The expression of 40 miRNAs was different between lymph node metastasis group and non-metastasis group.22 of them were up-regulated,and 18 of them were down-regulated.After clustering and PAM analysis and real-time RT-PCR technology demonstration,we screened out two kinds of tissue specific expression of miRNAs（miRNA-612 and miRNA-583）between lymph node metastasis group and non-metastasis group.miRNA-612 was obviously up-regulated and miRNA-583 obviously down-regulated.Conclusion The miRNAs detected in the miRNA microarray analysis may be characteristic molecular markers for lymph node metastasis of esophageal squamous cell carcinoma.And we can get better understanding of the miRNAs＇ functions in the development of esophageal squamous cell carcinomas,and go further in searching for relevant target molecule of clinical diagnosis and treatment in esophageal squamous cell carcinomas.

关 键 词：食管鳞状细胞癌 淋巴结转移 miRNA MIRNA芯片 实时定量RT-PCR 

分 类 号：R735.1[医药卫生—肿瘤]