应用微阵列芯片初步分析胃癌与癌旁组织miRNA差异表达谱

Preliminary analysis of miRNA expression profile of gastric cancer tissues by microarray

目的 应用微阵列芯片技术研究胃癌与癌旁正常组织miRNA差异表达谱,探讨miRNA与胃癌的发病关系.方法 提取5例胃癌患者癌组织及其对应癌旁正常组织中的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、癌旁正常组织进行差异基因筛选和聚类分析.采用实时定量PCR Taqman荧光探针法验证差异表达的hsa-miR-30c、hsa-miR-196a、hsa-miR-193b.利用在线靶基因预测软件对hsa-miR-30c的靶基因预测.结果 miRNA表达谱微阵列芯片筛选出显著差异表达的miRNA共16个,胃癌组织与癌旁正常组织比较,miRNA表达水平上调超过两倍且差异有统计学意义（P＜0.05）的有5个,分别是HS_100、hsa-miR-27a、hsa-miR-214＊、solexa-555-1991、hsa-miR-196,表达水平下调超过两倍且差异有统计学意义（P＜0.05）的有1 1个,分别是hsa-miR-502-3p、hsa-miR-29c＊、hsa-miR-64、hsa-miR-193b、hsa-miR-55 1b、hsa-miR-30c、hsa-miR-582-5p、hsa-miR-625＊、hsa-miR-660、hsa-miR-363、hsa-miR-486-5p.荧光定量PCR验证hsa-miR-30c,hsa-miR-196a和hsa-miR-193b表达与miRNA芯片的结果相符合.结论 胃癌与癌旁组织miRNA差异表达谱中的miRNA可能参与胃癌发生的调控,并将成为潜在的胃癌标志物.

Objective To analyze the expression profile of specific miRNA of gastric cancer tissues.Methods miRNA were isolated from five paired gastric cancer tissues and corresponding paracancerous normal tissues.To identify the profile of gastric cancer,miRNA hybridization and cluster analysis were processed by significance analysis of microarrays （SAM,version 2.1） and cluster software.The relative expression of hsa-miR-30c,hsa-miR-196a and hsa-miR-193b were validated by fluorescence Real-Time PCR using taqman probe.Further hsa-miR-30c taget gene was predictived by online software.Results Revealed by miRNA microarray detection,there were 16 miRNA with differential expression in gastric cancer tissues and para-cancerous normal tissues,among which HS_100,hsa-miR-27a,hsa-miR-214＊,solexa-555-1991 and hsa-miR-196 were significantly up-regulated,and hsa-miR-502-3p,hsa-miR-29c＊,hsa-miR-64,hsa-miR-193b,hsa-miR-551b,hsa-miR-30c,hsa-miR-582-5p,hsa-miR-625＊,hsa-miR-660,hsa-miR-363 and hsa-miR-486-5p were down-regulation.The relative expression of hsa-miR-30c,hsa-miR-196a and hsa-miR-193b was in high concordance with microarray.Conclusion There is specific miRNA expression profile in gastric cancer tissues,among which differentially-expressed miRNAs may be involved in the regulation of gastric cancer progress and serve as new potential biomarkers.