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目的:对miRNA-194在骨肉瘤中的作用机制进行探索和研究,为研究miRNA-194在骨肉瘤中的作用及可能的生物治疗提供理论依据和研究基础。方法:使用生物信息学软件对miRNA-194在骨肉瘤中可能的靶基因进行预测,并在基因水平和蛋白水平进行相应的验证。此外,利用荧光素酶基因报告实验对其靶基因(CDH2)进行DNA水平的直接验证。结果:使用生物信息学软件预测出了多个可能的潜在靶基因(367个,6014个和212个);荧光素酶基因报告实验显示野生PTEN-3’-UTR在miRNA-194上调组明显的低表达,同时下调组明显的高表达(P<0.05);同时,western实验显示在蛋白水平miRNA-194直接靶向作用于CDH2(P<0.05),而在m RNA水平则无明显作用(P>0.05)。结论:经预测和验证,CDH2是miRNA-194在骨肉瘤中的直接靶基因,为接下来关于miRNA-194在骨肉瘤中的各项研究奠定了良好的实验基础。
Objective: To explore the mechanism of miRNA-194 in osteosarcoma and to provide theoretical basis and research basis for studying the role of miRNA-194 in osteosarcoma and possible biological therapy. METHODS: Bioinformatics software was used to predict the possible target genes of miRNA-194 in osteosarcoma and to verify the gene and protein levels accordingly. In addition, luciferase gene reporter assay was used to directly verify the DNA level of its target gene (CDH2). Results: Multiple potential target genes (367, 6014 and 212) were predicted using bioinformatics software. The luciferase gene reporter assay showed that wild type PTEN-3’-UTR was significantly up-regulated in miRNA-194 (P <0.05). In the meantime, Western experiments showed that miRNA-194 directly targeted to CDH2 at protein level (P <0.05), while had no effect at m RNA level (P > 0.05). CONCLUSION: CDH2 is a direct target of miRNA-194 in osteosarcoma after the prediction and validation, which lays a solid foundation for further studies on miRNA-194 in osteosarcoma.