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目的 探索辐射诱导基因调控元件启动的造血生长因子表达及其对造血的保护作用。方法 本实验将携带Egr 1调控序列的FLT3配基 (FL)和EGFP基因双顺反子载体 (Egr EF)转染骨髓基质细胞系HFCL ,采用流式细胞仪、RT PCR、ELISA及细胞增殖法等观察细胞受照后Egr 1调控元件诱导的FL表达及促进造血细胞的增殖作用。 结果 在转染细胞HFCL/EF细胞中证实有外源性基因的整合和表达 ,在 2 5Gy辐射后 16h的细胞培养上清液中表明FL含量较照射前明显增高 ((P <0 0 1) ,) ;辐射后 10dHFCL/EF培养上清液对CD34+造血祖细胞的作用较辐射前具有明显的扩增作用 ((P <0 0 1) ,)。结论 在辐射后Egr 1启动子调控的FL基因表达明显增高并具有保护造血作用。
Objective To explore the expression of hematopoietic growth factors activated by radiation-induced gene regulatory elements and their protective effect on hematopoiesis. Methods FLT3 ligand (FL) and Egr EF carrying Egr 1 regulatory sequence were transfected into bone marrow stromal cell line HFCL by flow cytometry, RT-PCR, ELISA and cell proliferation assay Observed the expression of FL induced by Egr 1 regulatory elements after irradiation and promoted the proliferation of hematopoietic cells. Results The integration and expression of exogenous genes were confirmed in HFCL / EF transfected cells. The cell culture supernatant at 16h after 2 5Gy irradiation showed that the content of FL was significantly increased (P <0.01) (P <0.01)). The effect of the supernatant of HCFCL / EF culture on CD34 + hematopoietic progenitor cells 10 d after irradiation was significantly higher than that before radiation (P <0.01). Conclusion The expression of FL gene regulated by Egr 1 promoter after irradiation is significantly increased and has the effect of protecting hematopoiesis.