Involvement of NF-κB in the regulation of S100A6 gene expression in human hepatoblastoma cell line HepG2
Section snippets
Materials and methods
Cell cultures. Human liver cell lines, HepG2, PLC/ARF/5, Hep3B, and Chang were obtained from the American Type Culture Collection (Rockville, MD, USA). Cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (Life Technologies, Gaithersburg, MD, USA) at 37 °C in a 5% CO2 air environment.
Northern blot analysis and RT-PCR. HepG2 cells were stimulated with TNFα (0.5, 2, and 5 ng/ml, Sigma Chemical, St. Louis, MA, USA) for 24 h and total RNA was isolated from
S100A6 is differentially expressed in liver cell lines
To study the regulation of S100A6 gene in hepatocytes, we carried out Northern blot analysis on several liver cell lines cultured in DMEM with 10% FBS (Fig. 1). S100A6 mRNA was highly expressed in Chang and Hep3B cells but barely detected in HepG2 and PLC/ARF/5 cells. The expression levels of S100A6 in these cell lines were accordant with their cell growth rates; Chang and Hep3B had higher growth rate than other liver cell lines (data not shown). This result is in agreement with the previous
Acknowledgements
This study was supported in part by grants of Korea Ministry of Science and Technology (KGS2010212, KGS2110311, and NTM0020213).
References (26)
Functional roles of S100 proteins, calcium-binding proteins of the EF-hand type
Biochim. Biophys. Acta
(1999)S100: a multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles
Int. J. Biochem. Cell Biol.
(2001)- et al.
Molecular cloning of the cDNA for a growth factor-inducible gene with strong homology to S-100, a calcium-binding protein
J. Biol. Chem.
(1986) - et al.
Calcyclin as a marker of human epithelial cells and fibroblasts
Exp. Cell Res.
(1992) - et al.
Evidence that the time of entry into S is determined by events occurring in early G1
J. Biol. Chem.
(1988) - et al.
Identification of an AP-1-like sequence in the promoter region of calcyclin, a S-100-like gene. Enhancement of binding during retinoic acid-induced neuroblastoma cell differentiation
Neurosci. Lett.
(1994) - et al.
S100A6 protein as a marker for differential diagnosis of cholangiocarcinoma from hepatocellular carcinoma
Hepatol. Res.
(2002) The inducible transcription activator NF-kappa B: regulation by distinct protein subunits
Biochim. Biophys. Acta
(1991)- et al.
NF-kappaB as a frequent target for immunosuppressive and anti-inflammatory molecules
Adv. Immunol.
(1997) - et al.
Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction
Anal. Biochem.
(1987)