胃癌转移相关miRNA的筛选及生物信息学分析

Screening and bioinformatic analysis of microRNAs associated with metastasis of gastric cancer

目的:筛选与胃癌转移相关的微小RNA(microRNA,miRNA,miR),并进行生物信息学分析。方法:应用miRNA芯片对胃癌低转移细胞株(RF-1和MKN28)和高转移细胞株(RF-48、NCI-N87和KATOⅢ)进行miRNA差异表达谱分析,筛选差异表达的miRNA;应用实时荧光定量-PCR法对差异表达的miR-192、miR-215和miR-194进行验证,应用miRfocus在线数据库对筛选获得的miR-192、miR-193a、miR-194和miR-215进行靶基因预测及其信号转导通路进行分析。结果:与低转移细胞株比较,高转移细胞株中miR-192、miR-193a、miR-194和miR-215的表达水平明显上调,miR-105、miR-767、miR-149、miR-205、miR-30a、miR-30d、miR-365、miR-193b、miR-326、miR-100、miR-30b、miR-125b和miR-584的表达水平明显下调。实时荧光定量-PCR法对miR-192、miR-215和miR-194的表达进行验证,结果显示与miRNA芯片检测结果一致;生物信息学分析结果显示,miR-192、miR-193a、miR-194和miR-215及其靶基因参与肿瘤的发生、转移、细胞周期及范可尼贫血等通路。结论:胃癌高转移细胞中miR-192、miR-193a、miR-194和miR-215的上调可能与胃癌的发生和转移等有关。

Objective： To screen the microRNA （miRNA, miR） associated with metastasis of gastric cancer and to analyze its bioinformation. Methods： The differentially expressed miRNAs in gastric cancer cell lines with low metastatic potentials （RF-1 and MKN28） or high metastatic potentials （RF-48, NCI-N87 and KATO III） were analyzed and screened by miRNA microarrays. The differentially expressed miR-1 92, miR-21 5 and miR-194 were verified by real-time fluorescence quantitative-PCR. The targets prediction of miR-192, miR-193a, miR-194, miR-215 and signal pathway analysis were performed via miRfocus database. Results： As compared with the cells with low metastatic potentials, the expression levels of miR-192, miR-193a, miR-194 and miR-215 were up-regulated and the expression levels of miR-105, miR-767, miR-149, miR-205, miR-3Oa, miR-3Od, miR-365, miR-193b, miR-326, miR-lO0, miR-3Ob, miR-125b and miR-584 were down-regulated in highly metastatic cells. Up-regualtions of miR-192, miR-194 and miR-215 were validated by real-time fluorescence quantitative-PCR and the results were consistent with those of microRNA microarray. Bioinformatic analysis, functions of targets of miR-1 92, miR-193a, miR-194 and miR-215 were focused on the miRNA in cancer, pathway in cancer, cell cycle and Fanconi anemia pathway, etc. Conclusion= miR-192, miR-193a, miR-194 and miR-215 are overexpressed in highly metastatic gastric cancer cells, and they may be related to the progression and metastasis of gastric cancer.