| SUN Shi-long,DONG Juan-cong,JIN Shun-zi.Regulation of miR-503 on the expression of CD40 induced by irradiation in U937 cells[J].Chinese Journal of Radiological Medicine and Protection,2011,31(6):636-639 |
| Regulation of miR-503 on the expression of CD40 induced by irradiation in U937 cells |
| Received:April 23, 2011 |
| DOI：10.3760/cma.j.issn.0254-5098.2011.06.004 |
| KeyWords:miR-503 CD40 Irradiation Tumor Regulation |
| FundProject:国家自然科学基金面上项目(30870584);国家教育部留学回国人员启动基金项目(20101201) |
| Author Name | Affiliation | | SUN Shi-long | MH Radiobiology Research Unit, School of Public Health, Jilin University, Changchun 130021, China | | DONG Juan-cong | MH Radiobiology Research Unit, School of Public Health, Jilin University, Changchun 130021, China | | JIN Shun-zi | MH Radiobiology Research Unit, School of Public Health, Jilin University, Changchun 130021, China |
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| Abstract:: |
| Objective To study whether or not CD40 gene is a target of miR-503 in U937 cells, and to observe the regulatory effects of miR-503 on expression of CD40 induced by irradiation in U937cells. Methods The miR-503 sequence was inserted into pcDNA-DEST-47 plasmid to construct the eukaryotic expression vector (pcDNA-DEST-miR-503) and to construct the CD40 gene 3'-UTR luciferase reporter plasmid (psiCHECK2-CD40) at the same time. They were used to transfect U937 cells together for analysis of the regulatory effects of miR-503 on the expression of CD40. Meanwhile the miR-203 and miR-29b were used as controls to observe whether or not CD40 gene was a target of miR-503. The expression change of CD40 after irradiation, and the inhibitory effect of miR-503 on the expression of CD40 was confirmed by Western blot assay. The expression of miR-503 was detected by real-time RT-PCR (qPCR) after irradiation with doses of 5.0 Gy. Results The expression of luciferase in the group of transfected with pcDNA-DEST-miR-503 and psiCHECK2-CD40 plasmids was significantly lower than that in the group transfected with empty plasmid and CD40 gene (t=3.16, P<0.05). And the miR-203 and miR-29b could not inhibit the activity of CD40 luciferase, but the miR-503 could significantly inhibit the activity of it (t=5.25, P<0.01). The expression of CD40 protein in U937 cells was decreased after the cells were transfected with pcDNA-DEST-miR-503. After irradiation with dose of 5.0 Gy, the expression of miR-503 were increased (t=3.63-17.00, P<0.01) and the expression of CD40 protein decreased. Conclusions The CD40 gene might be the target of miR-503, and miR-503 could regulate the expression of CD40 gene.Irradiation could up-regulate the expression of miR-503 and inhibit the expression of CD40 protein in U937 cells. miR-503 might play a role in the process of regulation of irradiation on expression of CD40 protein in tumor cells. |
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