miR-15b靶基因预测、信号通路分析及靶基因鉴定

Prediction,pathway analysis and certification of miR-15b targeting genes

【目的】对miR-15b靶基因进行预测、信号通路分析及鉴定,以期为miR-15b的功能研究奠定基础.【方法】利用Target Scan Release 7.0,PicTar和PITA数据库对miR-15b靶基因进行预测,再用DAVID数据库对预测出的靶基因集进行功能富集分析(gene ontology-analysis)及信号转导通路富集分析(KEGG pathway analysis).随后利用双荧光素酶报告试验对预测出miR-15b的靶基因丙酮酸脱氢酶激酶4(PDK4)和CD28进行验证.【结果】3个软件共同预测出miR-15b的靶基因有305个,其靶基因集合功能富集于蛋白激酶活性、GTP结合蛋白调控等分子功能,蛋白氨基酸磷酸化、细胞周期调控等生物学过程及微管相关复合体、转录因子复合体等细胞组分上.信号转导通路则显著富集于癌症相关信号通路.双荧光素酶报告实验结果显示CD28为miR-15b的靶基因,而PDK4不是miR-15b的靶基因.【结论】miR-15b在细胞增殖、凋亡和细胞周期调控等过程中发挥重要调控作用,且miR-15b通过调控CD28的表达对T细胞的受体激活发挥调控作用.

【Objective】To predict,analyze and certificate target genes of miR-15 bwith the aim of providing certain basis for the functional research of miR-15 b.【Method】The target genes of miRNA-15 bwere screened through Target Scan Release 7.0,PicTar and PITA databases.Subsequently,bioinformatic analysis of these target genes were performed by Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes Pathway analysis（KEGG Pathway analysis）by DAVID database.Dual Luciferase Reporter gene assays were used to certificate.【Result】There were 305 target genes were predicted by the three databases,and the gene set mainly located in protein kinase activity,purine nucleoside binding（GO molecular function）,protein amino acid phosphorylation,regulation of cell cycle（GO biology process）and microtubule associated complex,transcription factor complex（GO cellular component）.The KEGG Pathway analysis demonstrated that the gene set mostly was located in Pathways in cancer.The result of Dual-luciferase re-port experiment showed that CD28,but PDK4 was the target gene of miR-15 b.【Conclusion】miR-15 b plays a significant role in cell proliferation,cell apoptosis and cell cycle regulation,which can regulate coreceptor activity of T cells by targeting CD28.