| DU Nan,PEI Xue-tao,ZHOU Jin-ming,SUN Jun-zhong,FU Yan,ZHAO Hui.Egr-1 promoter regulating effect on granulocyte-macrophage colony-stimulating factor (GM-CSF) gene expression induced by doxorubicin and ionizing radiation[J].Chinese Journal of Radiological Medicine and Protection,2009,29(3):249-252 |
| Egr-1 promoter regulating effect on granulocyte-macrophage colony-stimulating factor (GM-CSF) gene expression induced by doxorubicin and ionizing radiation |
| Received:June 26, 2008 |
| DOI：10.3760/cma.j.issn.0254-5098.2009.03.001 |
| KeyWords:Early growth response-1 Ionizing radiation Doxorubicin Granulocyte-macrophage colony-stimulating factor Radical oxygen intermediates |
| FundProject:国家自然科学基金(30572144);解放军总医院第一附属医院新技术重大项目(ZD200502) |
| Author Name | Affiliation | | DU Nan | Department of Oncology and Hematology, The First Affiliated Hospital, Chinese PLA General Hospital, Beijing 100037, China | | PEI Xue-tao | 军事医学科 | | ZHOU Jin-ming | 学院九所 | | SUN Jun-zhong | Department of Oncology and Hematology, The First Affiliated Hospital, Chinese PLA General Hospital, Beijing 100037, China | | FU Yan | Department of Oncology and Hematology, The First Affiliated Hospital, Chinese PLA General Hospital, Beijing 100037, China | | ZHAO Hui | Department of Oncology and Hematology, The First Affiliated Hospital, Chinese PLA General Hospital, Beijing 100037, China |
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| Abstract:: |
| Objective To explore the regulating effects of Egr-1 promoter activated by ionizing radiation (IR) and doxorubicin (ADM) on the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) genes.Methods The human GM-CSF cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with IRES(internal ribosome entry site) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter(Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by liposome transfection. And the cells were exposure to ADM and IR. The activity of EGFP in HFCL/EG cells were detected by FACS. The effect of N-acetylcysteine on the expression of EGFP following exposure to ADM and IR was examined. The amounts of GM-CSF in HFCL/EG after chemotherapy or radiation were measured with ELISA. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood were also studied. RT-PCR analysis for the expression of GM-CSF mRNA in HFCL/EG after exposure to ADM or IR.Results The percentage of EGF HFCL/EG cells exposed to ADM and IR was increased compared with non-treatment group (1.2% and 15.2% vs 18.2%, t=5.11,P<0.01). The levels of secreted GM-CSF in HFCL/EG cells exposed to ADM and IR was increased (P<0.01), but no difference between ADM group and IR group (P>0.05). The expression of EGFP by HFCL/EG treated with ADM and IR was significantly decreased by N-acetylcysteine. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM in ADM group and IR group were significantly higher than that in HFCL group and non-treatment group. However, The CFU-GM count of IR group was higher than that of ADM group. The expression of GM-CSF mRNA in HFCL/EG cells exposed to ADM and IR was significantly increased(t=4.37,P<0.01).Conclusions GM-CSF gene expression regulated by Egr-1 promoter induced by ADM and IR could help the recovery from hematopoietic injury. |
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