| 杜香 王飞 韩康 祁健 赵廷宝△.miRNA-194 在骨肉瘤中靶基因的预测和验证[J].,2016,16(18):3581-3585 |
| miRNA-194 在骨肉瘤中靶基因的预测和验证 |
| The Prediction and Verification of Target Genes of miRNA-194 inOsteosarcoma |
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| DOI： |
| 中文关键词: miRNA-194 CDH2 机制 骨肉瘤 |
| 英文关键词: MicroRNA-194 CDH2 Mechanism Osteosarcoma |
| 基金项目:国家自然科学基金项目(31500856) |
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| 中文摘要: |
| 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mRNA 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。 |
| 英文摘要: |
| To forecast and verify the target genes of miRNA-194 in human osteosarcoma and establish some
theoretical basis for osteosarcoma treatment and element experience of miRNA-194 for further research. Potential target gene
of miRNA-194 was predicted using bioinformatics and were verified in gene and protein levels. Moreover, luciferase reporter assay was
used to confirm whether CDH2 was direct targets of miRNA-194. Potential target genes of miRNA-194 were predicted by using
bioinformatics. Moreover, luciferase activity of the wild-type PTEN-3'-UTR reporter was significantly suppressed in miRNA-194
up-regulation group compared with other groups in SOSP-9607 cells (p<0.01) while down-regulation group could significantly increase it
(p<0.01). At the same time, western blot analysis revealed that miRNA-194 expression could affect the expression of CDH2 protein while
there was no significant difference on mRNA level (P>0.05). CDH2, one target gene of miRNA-194, was predicted and
verified, which provided the theoretical basis for the next study of miRNA-194 in the osteosarcoma. |
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