| Zhang Haiqin,Dong Juancong,Gao Hui,et al.Target regulation of miR-9 to the expression of NRP1 and its role in radiation effects[J].Chinese Journal of Radiological Medicine and Protection,2014,34(10):725-728 |
| Target regulation of miR-9 to the expression of NRP1 and its role in radiation effects |
| Received:May 03, 2014 |
| DOI：10.3760/cma.j.issn.0254-5098.2014.10.002 |
| KeyWords:miR-9 NRP1 A549 Targeted regulation Ionizing radiation |
| FundProject:国家自然科学基金（30870584，81371890）；教育部高等学校博士学科点专项科研基金（20120061110063） |
| Author Name | Affiliation | E-mail | | Zhang Haiqin | College of Public Medicine, Key Laboratory of Radiobiology, Ministry of Health, Jilin University, Changchun 130021, China | | | Dong Juancong | College of Public Medicine, Key Laboratory of Radiobiology, Ministry of Health, Jilin University, Changchun 130021, China | | | Gao Hui | College of Public Medicine, Key Laboratory of Radiobiology, Ministry of Health, Jilin University, Changchun 130021, China | | | Zuo Siyao | College of Public Medicine, Key Laboratory of Radiobiology, Ministry of Health, Jilin University, Changchun 130021, China | | | Jin Linlin | College of Public Medicine, Key Laboratory of Radiobiology, Ministry of Health, Jilin University, Changchun 130021, China | | | Liu Libo | College of Public Medicine, Key Laboratory of Radiobiology, Ministry of Health, Jilin University, Changchun 130021, China | liulb@jlu.edu.cn | | Jin Shunzi | College of Public Medicine, Key Laboratory of Radiobiology, Ministry of Health, Jilin University, Changchun 130021, China | |
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| Abstract:: |
| Objective: To explore the effect of miR-9 on the expression of NRP1 and its radiation effects in A549 cells. Methods: Bioinformatics was used to analyze the potential binding sites of has-miR-9 and NRP1-3'UTR. The miR-9 sequence was inserted into pcDNA-DEST-47 plasmid to construct the eukaryotic expression vector (pcDNA-DEST-miR-9) and to construct the NRP1 gene 3'UTR luciferase reporter plasmid (pEZX-MT05) at the same time. They were simultaneously transferred into A549 cells for analysis of the regulatory effect of miR-9 on the expression of NRP1. Meanwhile miR-29b was used as a negative control to observe whether or not NRP1 gene was a target of miR-9. After 10 Gy irradiation, the expression of NRP1, and the inhibitory effect of miR-9 on it was confirmed by Western blot assay. The expression of miR-9 was detected by real-time PCR. Results: It was found that miR-9 reduced the luciferase activity of NRP1-3'UTR wild plasmid (t=3.906, P<0.05) but not NRP1-3'UTR mutant plasmid. This luciferase activity was not inhibited by other types of miRNA (miR-29b). The expression of NRP1 protein in A549 cells was decreased after the cells were transfected with miR-9 mimic. After irradiation with dose of 10 Gy, the expression of miR-9 were decreased (t=37.319, P<0.05) and the expression of NRP1 protein were increased. Conclusions: miR-9 regulates the expression of NRP1 by targeting 3'UTR site of NRP1 gene in A549 cells. |
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