miRNA-194 在骨肉瘤中靶基因的预测和验证 |
| |
引用本文: | 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].现代生物医学进展,2016,16(18):3581-3585. |
| |
作者姓名: | 杜香 王飞 韩康 祁健 赵廷宝 |
| |
作者单位: | 济南军区总医院脊髓修复科 |
| |
基金项目: | 国家自然科学基金项目(31500856) |
| |
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(P0.05);同时,western实验显示在蛋白水平miRNA-194直接靶向作用于CDH2(P0.05),而在m RNA水平则无明显作用(P0.05)。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。
|
关 键 词: | miRNA-194 CDH2 机制 骨肉瘤 |
The Prediction and Verification of Target Genes of miRNA-194 in
Osteosarcoma |
| |
Abstract: | 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. |
| |
Keywords: | MicroRNA-194 CDH2 Mechanism Osteosarcoma |
本文献已被 CNKI 等数据库收录! |
| 点击此处可从《现代生物医学进展》浏览原始摘要信息 |
| 点击此处可从《现代生物医学进展》下载免费的PDF全文 |
|