| DU Nan,PEI Xuetao,LUO Chengji.Studies on the expression of hematopoietic growth factors regulated by Egr-1 promoter[J].Chinese Journal of Radiological Medicine and Protection,2000,20(5):306-309 |
| Studies on the expression of hematopoietic growth factors regulated by Egr-1 promoter |
| Received:October 09, 1999 |
| DOI： |
| KeyWords:Radiation Egr-1 Promoter FLT3 ligand Gene therapy |
| FundProject:国家自然科学基金资助项目(39900040);国家杰出青年自然科学基金资助项目(39825111) |
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| Abstract:: |
| Objective To explore the regulatory effects of radiation-inducible gene on the expression of FLT3 ligand genes. Methods The human EL cDNA and EGFP cDNA were linked together with IRES and then inserted into the expression vector pCI-Egr-1, which was constructed by substituting CMV promoter in pCI-neo with the Egr-1 promoter(Egr-EF).The vector was transferred into human bone marrow stromal cell line HFCL by lipofectin TM .The cells were exposed to γ-radiation from 60 Co source at 0.5-20Gy.The contents of green fluorescence in HFCL/EF cells were detected with FACS.The expression of FL in HFCL/EF cells was confirmed with RT-PCR and ELISA respectively.The effects of FL in HFCL/EF cultural supernatants on expansion of CD34 + cells were also studied. Results The amounts of secreted FL in serum-free supernatants of Egr-EF were significantly higher than in the control group.EGFP and FL cDNA were successfully integrated and expressed in the cells,which were confirmed by FACS,RT-PCR and ELISA analysis.On day 10 of culture the number of CD34+ cells was significantly higher than that of the non-radiation group. Conclusion These in vitro data provide an experimental basis for the in vivo use of gene therapy of hematopoietic growth factor gene regulated by Egr-1 promoter to protect hematopoiesis from radiation injury. |
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