鹿茸顶端组织miRNA芯片制备及差异表达miRNA的筛选  

Preparation of MicroRNA Chip from Deer Antler Tip Tissue and Screening of Differential Expression MicroRNAs

作　　者：崔东明[1] 孟星宇[1] 李婷[1] 田玉华[1] 李沐[1] 李璐[1] 胡薇[1] 

机构地区：[1]吉林农业大学生命科学学院,长春130118

出　　处：《经济动物学报》2016年第2期83-88,共6页Journal of Economic Animal

基　　金：国家自然科学基金项目(31572372;30972083);吉林省科技发展计划项目(20090574)

摘　　要：为制备梅花鹿鹿茸顶端间充质组织及软骨组织miRNA芯片,分离鹿茸间充质和软骨组织细胞,利用TRIzol试剂法分别提取细胞的总RNA,并进行miRNA生物素标记,将标记的小RNA在miRNA芯片上进行杂交反应,再进行芯片扫描及数据处理,应用相对荧光定量Real-time PCR法验证部分miRNA芯片结果的可靠性。研究结果显示:筛选得到鹿茸间充质和软骨组织miRNA差异表达谱。间充质部位存在的miRNA为289个,软骨部位存在的miRNA条数为304条。其中间充质组织中117个miRNA表达上调,71个miRNA表达下调;软骨组织中97个miRNA表达上调,87个miRNA表达下调;两个组织中共同存在且具有显著差异性表达的miRNA为158个。Real-time PCR法验证结果显示,相对于间充质细胞,miR-1、miR-18a、miR-18b、miR-19a、miR-19b、miR-20b和miR-let-7f的表达均上调;而miR-99b、miR-130b及miR-184的表达均下调,与芯片结果一致。To prepare miRNA chip of sika deer antler tissue, mesenchymal and cartilage tissue were separated from deer antler tip. Total RNA was extracted with TRIzol reagent and hybridization reac- tion was conducted. Afterwards, chip scanning and data processing were conducted. Finally, partial results of miRNA chips were verified by Real-time PCR. The results showed that there were 289 miRNAs at mesenchymal part and 304 miRNAs at cartilage part. Expressions of 117 miRNAs in me- senehyma were up-regulated and those of 71 miRNAs were down-regulated. Expressions of 97 miR- NAs in cartilage tissue were up-regulated and those of 87 miRNAs were down-regulated. There were 158 miRNAs with significantly differential expression in two tissues. Real-time PCR confirmed that expressions of miR-19a, miR-20b and miR-let-7f were up-regulated compared with mesenchymal cell. However, expressions of miR-99b and miR-184 were down-regulated, which was consistent with result of chip.

关 键 词：鹿茸 间充质 软骨 MIRNA芯片 差异表达 

分 类 号：S852.23[农业科学—基础兽医学]