技术与方法 |
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Study on A GFP Expression System for Detecting Estrogenic Compounds |
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Abstract Based on transcriptional regulation of estrogenic compounds, a recombinant 3×ERE EGFP-N1 reporter vector was constructed by inserting 3×ERE TATA sequence and transfected into MCF-7 breast cancer cells. Stable transfectants were selected and their GFP fluorescence intensity was measured using flow cytometry after adding E2 and other chemicals. 17β-estradiol(E2) induced a dose-dependent increase in GFP intensity in the cells, reaching maximum response at 1×10-10 mol/L and EC50 value was 1.5×10-11 mol/L, so done by phytoestrogen daidzein and polydatin , their EC50 values were 2.4×10-7 mol/L and 6.2×10-6 mol/L respectively, but grape seed polyphenols (GSP) had no significant estrogenic activity. Estrogen antagonist tamoxifen blocked GFP expression induced by E2 in a dose-dependent way and had a maximum inhibition response at 1×10-7 mol/L. The cell model can be used to detect ligands of estrogen receptor by testing GFP intensity induced by chemicals
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Received: 29 May 2006
Published: 25 October 2006
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