The Study Of The Effect And Mechanism Of Daxx Regulates Cholesterol Contents In HepG₂Cells

Object: To observe the effects and mechanisms of the functional domain and thefull-length of Daxx on the celluar cholesterol. And to investigate a new way for theprevention and treatment of hyperlipidemia and related acute coronary events.Methods: The Fas death domain-associated protein (Daxx) functional domain mutantand full-length Daxx eukaryotic expression vectors were stably transfected intoHepG2cells. Whether to establish stable cell lines successfully was identified byWestern Blotting. The cholesterol contents in intracellular were evaluated by enzymefluorescence and HPLC. The interaction of Daxx and AR was observed by Confocal.The mRNA and protein expression of SCAP and SREBP in cells were detected byRT-PCR and Western Blot respectively.Results: In the pCDNA3.1(+)/Daxx/DM (626-740) or pCDNA3.1(+)/Daxxgroups, the cholesterol contents were decreased which was analyed by enzymefluorescence and HPLC. The expression of Daxx and AR in the cytoplasm and thenucleus found by confocal was strengthened; RT-PCR and Western Blot demonstratedthat the mRNA and protein of SCAP and SREBP were decreased.Conclusion: Daxx reduced the expression of SCAP by combinding of AR, and theninhibited the activation of SREBPs, thus decreaced down the cholesterol contents ofliver cells.