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MI(N28)和高转移细胞株(RF-48、NCI—N87和勋打oIII)进行miIWA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-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.