| Background: Hypercholesterolemia is one of the serious risk factors ofthe formation and development of atherosclerotic plaque, as an importantnuclear transcription factor of cholesterol synthesis related genes,Androgen receptor was confirmed probably loss it’s transcriptionalactivation function through interaction with DAXX, but the specificmolecular mechanism remains to be further validation.Objective: To Confirm the interaction of DAXX and AR in HepG2cellsand suppress the transcriptional regulation to cholesterol synthesis relatedgene.Method: HepG2cells was stable transfected with pCDNA3.1(+)/DAXX,then Co-immunoprecipitation(IP) was recommended to make sure theinteraction between DAXX and AR; Oil Red Staining, Enzyme couplingcolorimetric and High performance liquid chromatography were adoptedto detect the content of total cholesterol in cells; Western Blotting andRT-qPCR were applied to measure the expression level of AR, SREBP-2and HMGCR; Use the TP with final concentration of1μmol/L as apositive drug to confirm that DAXX could competitively inhibited thetranscriptional activation of AR; siRNA was employed to exhaust the endogenous DAXX for detecting the expression of AR,SREBP-2andHMGCR.Result: DAXX could combined with AR in HepG2cells, but does notaffect the transcription and expression of AR;The high expressionof exogenous DAXX could significantly reduce the levels of totalcholesterol in HepG2cells, and seriously inhibit the expressionof SREBP-2and HMGCR intracellular; DAXX could effectively repressthe acceleration of testosterone propionate to cholesterol synthesis, andaccompanied by distinctly suppressed of the expression of SREBP-2andHMGCR; After depletion of endogenous DAXX, the levels of SREBP-2and HMGCR protein and mRNA were obviously restored in HepG2cells.Conclusion: DAXX is capable of combining to AR in HepG2cells, andthis function would suppress the biosynthesis of cholesterol throughdown-regulate the expression of SREBP-2and HMGCR. |
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