title: Genistein upregulates LDLR levels via JNK-mediated activation of SREBP-2.
abstract: BACKGROUND
  Genistein has been proved in vitro and in vivo to lower LDLR level. It is also widely consumed and implicated for its anti-atherogenic effects. However, the molecular mechanism by which genistein lowers the LDL level is still unknown.
  
  
  OBJECTIVE
  To understand the anti-atherogenic molecular mechanism of action, genistein was investigated for its impact on the expression of LDLR, the receptor for LDL cholesterol, and related signaling pathways in a human hepatoma cell line.
  
  
  METHODS
  HepG2 cell was used for the experiments. Genistein with different concentrations was diluted in media and was incubated for 24 h or more as indicated. Protein levels were measured by western blotting, and mRNA expression was detected by RT-qPCR. Chromatin immunoprecipitation assay (CHIP) assay was used to determine protein binding levels, and luciferase assay was used to measure promoter activity.
  
  
  RESULTS
  Genistein increased the mRNA and protein levels of LDLR in a time-dependent manner. Genistein increased the transcriptional activity of the LDLR promoter containing the reporter gene (pLDLR-luc, -805 to +50). But the sterol regulatory element deletion mutant construct failed to be activated by genistein. Genistein increased the nuclear fraction of SREBP-2 and the DNA-binding activity of SREBP-2 to LDLR promoter, as assessed by CHIP. The genistein-phosphorylated JNK inhibitor (SP600126) abolished the genistein-stimulated levels of LDLR and the nuclear SREBP-2. The addition of cholesterol up to 5 µg/mL for 24 h did not affect the effect of genistein on LDLR protein expression. Even the addition of 40 µM genistein increased the cholesterol uptake by more than 10% in the human hepatoma cell line.
  
  
  CONCLUSIONS
  Our data support the idea that genistein may have anti-atherogenic effects by activating JNK signals and SREBP-2 processing, which is followed by the upregulation of LDLR.
authors.affiliation: Department of Food and Nutrition, College of Human Ecology, Yonsei University, Seoul, South Korea.
authors.forename: Medicia
authors.initials: M
authors.lastname: Kartawijaya
authors.affiliation: Department of Food and Nutrition, College of Human Ecology, Yonsei University, Seoul, South Korea.
authors.forename: Hye Won
authors.initials: HW
authors.lastname: Han
authors.affiliation: Department of Food and Nutrition, College of Human Ecology, Yonsei University, Seoul, South Korea.
authors.forename: Yunhye
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Food and Nutrition, College of Human Ecology, Yonsei University, Seoul, South Korea; leeseungmin@yonsei.ac.kr.
authors.forename: Seung-Min
authors.initials: SM
authors.lastname: Lee
keywords: JNK; LDL receptor; SREBP-2; cholesterol; genistein