Regulation Of ETS And C/EBP On Apolipoprotein F

Objective: apolipoprotein F(ApoF) is found in recent years, theplasmaconcentration are closely associated with dyslipidemia. In thissubject we investigated the effect of ETS and C/EBP on apolipoproteinF expression.Methods:1. ApoF promoter region is build on PGL4vector2. Through pumbed genedatabase the potential transcription factor expression of the boxthat this topic havepreviously predicted is designed, and transcription factor expression cloning is builtby the method of gene synthesis method3. transcription factors and theircorresponding dominant deactivation mutant with the promoter are Co-transfection4.the interaction between cis-acting elements and trans-acting factors is test byelectrophoretic mobility of the detection and chromatin immune coprecipitation5.theprotein and mRNA expression level of apoF and relevant transcription factor thatcome from human sources of different tissue cell are test by the westernblot andrealtime PCR6.the protein level of C/EBP alpha and C/EBP beta in liver tissue of celllines is test by the westernblot7. whether the ETS-1、ETS-2、C/EBP alpha、ETS–1and C/EBP alpha、 ETS-2and C/EBP alpha promot the ApoF are test by thewesternblot8. the interactions among transcription factors are test by immunecoprecipitationResults:1.PGL4-ApoF198fluorescent reporter gene is successfullyconstructed;2C/EBP and HNF3family are successful build in pcDNA3.1expression vector;3.the protein and mRNA expression level of apoF in liver tissue ishigher than other cell;4.Overexpression of HNF3have no significant activation onapolipoprotein F promoter activities;5.Overexpression of C/EBP enhanceapolipoprotein F promoter activities, however,overexpression of C/EBP dominantinactivation mutant inhibit apolipoprotein F promoter activities;6.the protein expression level of E TS1、 ETS2、 C/EBPα及C/EBPβin liver tissue are higherthan other cell;7. the ETS-1、ETS-2、C/EBP alpha、ETS–1and C/EBP alpha、ETS-2and C/EBP alpha can promot the ApoF; ETS-1and C/EBP alpha have synergy to ApoF8.there are interaction between ETS-1and C/EppsConclusion:1. C/EBP alpha transcription factor directly combine ApoF promoter,regulate the transcription of ApoF.2. ETS-1has interaction between C/EBP alpha,have synergy to ApoF.