A Combination of Transcriptomics and Metabolomics Uncovers Enhanced Bile Acid Biosynthesis in HepG2 Cells Expressing CCAAT/Enhancer-Binding Protein 尾 (C/EBP尾), Hepatocyte Nuclear Factor 4伪 (HNF4伪), an
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- 作者：Marina Blazquez ; Aitor Carretero ; James K. Ellis ; Toby J. Athersuch ; Rachel Cavill ; Timothy M. D. Ebbels ; Hector C. Keun ; Jos茅 V. Castell ; Agust铆n Lahoz ; Roque Bort
- 刊名：Journal of Proteome Research
- 出版年：2013
- 出版时间：June 7, 2013
- 年：2013
- 卷：12
- 期：6
- 页码：2732-2741
- 全文大小：548K
- 年卷期：v.12,no.6(June 7, 2013)
- ISSN：1535-3907
文摘
The development of hepatoma-based in vitro models to study hepatocyte physiology is an invaluable tool for both industry and academia. Here, we develop an in vitro model based on the HepG2 cell line that produces chenodeoxycholic acid, the main bile acid in humans, in amounts comparable to human hepatocytes. A combination of adenoviral transfections for CCAAT/enhancer-binding protein 尾 (C/EBP尾), hepatocyte nuclear factor 4伪 (HNF4伪), and constitutive androstane receptor (CAR) decreased intracellular glutamate, succinate, leucine, and valine levels in HepG2 cells, suggestive of a switch to catabolism to increase lipogenic acetyl CoA and increased anaplerosis to replenish the tricarboxylic acid cycle. Transcripts of key genes involved in bile acid synthesis were significantly induced by approximately 160-fold. Consistently, chenodeoxycholic acid production rate was increased by more than 20-fold. Comparison between mRNA and bile acid levels suggest that 12-alpha hydroxylation of 7-alpha-hydroxy-4-cholesten-3-one is the limiting step in cholic acid synthesis in HepG2 cells. These data reveal that introduction of three hepatocyte-related transcription factors enhance anabolic reactions in HepG2 cells and provide a suitable model to study bile acid biosynthesis under pathophysiological conditions.