Abstract:
:The human liver fatty acid binding protein (hFABP1) participates in cellular long-chain fatty acid trafficking and regulation of lipid metabolism and changes in hFABP1 are associated with an increased risk for type 2 diabetes, cardiovascular disease (CVD), and metabolic syndromes. Gene regulation of hFABP1 is not fully understood. Therefore, in the present study, the full length hFABP1 promoter (nucleotides -2125 to +51) and a series of truncated promoter regions were cloned. A luciferase reporter assay revealed that nucleotides -255 to +50 in the promoter region contained full of maximum hFABP1 promoter activity compared with the full length promoter. Furthermore high activity was shown when the plasmid was transfected into liver-derived cells such as the human hepatoblastoma cell line HepG2 and the hepatoma cell line Huh7. TFSEARCH and TESS programs were used to predict potential transcription factor binding sites. Two putative binding sites for the liver-enriched transcription factors hepatocyte nuclear factor 3β (HNF3β) and CCAAT/enhancer binding protein α (C/EBPα) were identified in the -255 nt to -155 nt hFABP1 promoter region. Site-directed mutagenesis of these two sites reduced dramatically hFABP1 promoter activity. In addition, the electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP) revealed that these binding sites were recognized by HNF3β and C/EBPα respectively. Overexpression of HNF3β and C/EBPα enhanced the transcription of hFABP1 and consequently improved the protein level of hFABP1 in HepG2 cells, while knockdown of HNF3β and C/EBPα showed the inverse effects. Taken together, the hFABP1 gene is highly transcribed in liver-derived cells, and regulated predominantly by liver-enriched transcription factors HNF3β and C/EBPα.
journal_name
Biochimiejournal_title
Biochimieauthors
Wu YL,Peng XE,Wang D,Chen WN,Lin Xdoi
10.1016/j.biochi.2011.08.006subject
Has Abstractpub_date
2012-02-01 00:00:00pages
384-92issue
2eissn
0300-9084issn
1638-6183pii
S0300-9084(11)00311-7journal_volume
94pub_type
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