Identification of intracellular pathways through which TGF-β1 upregulates URG-4/URGCP gene expression in hepatoma cells

Life Sci. 2016 Jan 1:144:121-8. doi: 10.1016/j.lfs.2015.12.010. Epub 2015 Dec 4.

Abstract

Aims: Up-regulated gene 4 (URG-4/URGCP) was strongly expressed in Hepatitis B infected liver and correlated with HBxAG (Hepatitis B x Antigen) protein and found to promote hepatocellular cancer. Transforming growth factor (TGF-β1) is a multifunctional protein that effects cell proliferation, growth inhibition, differentiation and other functions. However, the mechanism of URG-4/URGCP regulation by TGF-β1 and its significance in cancer progression remains largely unknown.

Main methods: The effect of TGF-β1 on URG-4/URGCP gene was determined using REAL TIME PCR at mRNA level and Western blotting/immunofluorescence at protein level. Transient transfection assays were carried out to find out which site of promoter is upregulated by TGF-β1.

Key findings: We report the upregulation of URG-4/URGCP gene expression by TGF-β1 in hepatoma cells along with prostate cancer cells, PC3. Transient transfection assays showed that the -109 to +63 promoter region contained the minimal TGF-β1 response elements. TGF-β1 markedly stimulated the URG-4/URGCP mRNA and protein that was blocked by MEK1 [MAPK (Mitogen-Activated Protein Kinase)/ERK (extracellular signal-regulated kinase) kinase 1] inhibitor, PD98059 and PI3K inhibitor, wortmannin.

Significance: These studies show for the first time that TGF-β1 upregulates the expression of URG-4/URGCP in human hepatocytes and identifies the signaling pathways underlying this response.

Keywords: MAPK/ERK; PI3K; Promoter; TGF-β; URG-4/URGCP; Upregulation.

MeSH terms

  • Androstadienes / pharmacology
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Disease Progression
  • Enzyme Inhibitors / pharmacology
  • Flavonoids / pharmacology
  • Gene Expression / drug effects
  • Humans
  • MAP Kinase Kinase 1 / antagonists & inhibitors
  • MAP Kinase Kinase 1 / biosynthesis
  • MAP Kinase Kinase 1 / genetics
  • MAP Kinase Signaling System / drug effects
  • Male
  • Neoplasm Proteins / biosynthesis*
  • Neoplasm Proteins / genetics
  • RNA, Messenger / biosynthesis
  • Transfection
  • Transforming Growth Factor beta1 / pharmacology*
  • Up-Regulation / drug effects
  • Wortmannin

Substances

  • Androstadienes
  • Enzyme Inhibitors
  • Flavonoids
  • Neoplasm Proteins
  • RNA, Messenger
  • Transforming Growth Factor beta1
  • URGCP protein, human
  • MAP Kinase Kinase 1
  • MAP2K1 protein, human
  • 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
  • Wortmannin