Studies on the expression of hematopoietic growth factor modulated by Egr-1 promoter sequence in transcriptional targeting by chemotherapeutic agents
- VernacularTitle:化疗诱导Egr-1启动子调控造血因子基因表达的实验研究
- Author:
Nan DU
;
Xuetao PEI
;
Jinming ZHOU
- Publication Type:Journal Article
- Keywords:
Egr-1 protein;
granulocyte-macrophage colony-stimulating factor;
hematopoietic stem cells;
reactive oxygen species
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the chemotherapeutic agents-induced modulating effects of Egr-1 promoter sequence on GM-CSF expression and its protective effect against injury to haematopoiesis due to chemotherapy.Methods Human GM-CSF cDNA and enhanced green fluorescent protein(EGFP) cDNA were linked to internal ribosome entry site(IRES) respectively,and then recombined to pCIneo vector containing Egr-1 promoter(Egr-EG).The vector was transferred into human bone marrow stromal cell line HFCL by lipofection,and the HFCL/EG cells were then finally constructed.MTT assay was performed to determine the effects of cisplatin(DDP),5-fluorouracil(5-FU),gemcitabine(GEM) and paclitaxel(PTX) on the survival rates of HFCL/EG and HFCL cells,and the IC50 of such agents against HFCL/EG and HFCL cells were calculated.The percentage of HFCL/EG cells which positively expressed EGFP was assessed by both flow cytometry and inverted fluorescence microscope.The effects of the active oxygen inhibitor N-acetylcysteine(NAC) on the expression of GM-CSF,which was modulated by promotor Egr-1,were detected by ELISA.The effects of HFCL/EG supernatant on CFU-GM after being exposed to chemotherapeutic agents were examined.Results With different sensitivity to DDP,5-FU,GEM and PTX,HFCL/EG cells were successfully constructed.The drugs showed higher IC50 value against HFCL/EG cells than HFCL cells(P
