Korean J Hepatol.  2009 Dec;15(4):486-495. 10.3350/kjhep.2009.15.4.486.

Transcriptional profiling and Wnt signaling activation in proliferation of human hepatic stellate cells induced by PDGF-BB

• Shin H
• Park SY
• Lee KB
• Shin E
• Nam SW
• Lee JY
• Jang JJ

• KMID: 1384056
• DOI: http://doi.org/10.3350/kjhep.2009.15.4.486

Abstract

BACKGROUND/AIMS
This study aimed to better understand gene expression profiles of human hepatic stellate cell (HSC) activation and the relationship with the Wnt signaling pathway. METHODS: The global transcript levels in platelet derived growth factor-BB (PDGF-BB)-stimulated hTERT HSCs were analyzed using oligonucleotide microarrays. Oligonucleotide microarrays with 19K human oligo chips were performed to obtain gene expression profiles associated with proliferation in human hTERT HSCs. The microarray data was verified by real time quantitative PCR and expression of the components of Wnt signaling was analyzed by Western blot. RESULTS: Microarray data showed 243 up-regulated and 265 down-regulated genes in PDGF-BB-treated HSCs. The changes in expression of glypican3 and BH3 interacting domain death agonist (BID) mRNA in real time quantitative PCR, especially among the highly up- or down-regulated genes, were statistically consistent with the microarray data. The Wnt signaling pathway components, frizzled10 (FZD10) and calcium/calmodulin-dependent protein kinase II alpha (CAMK2A), showed increased expression in the short time course microarray and the up-regulation of FZD10 also occurred at the protein level. Our data showed various gene expression profiles during activation of human HSC. CONCLUSIONS: The up-regulated expression of FZD10 and CAMK2A suggests that the Wnt/Ca2+ signaling pathway is active in hTERT HSCs and may participate in HSC activation and proliferation

Keyword

Hepatic stellate cells; PDGF-BB; Proliferation; Oligonucleotide microarray; Wnt signaling

MeSH Terms

Angiogenesis Inducing Agents/*pharmacology
Blotting, Western
Calcium-Calmodulin-Dependent Protein Kinase Type 2/genetics/metabolism
Cell Line
Cell Proliferation
Frizzled Receptors/genetics/metabolism
Gene Expression Profiling
Hepatic Stellate Cells/cytology/*metabolism
Humans
Oligonucleotide Array Sequence Analysis
Platelet-Derived Growth Factor/*pharmacology
Polymerase Chain Reaction
Receptors, G-Protein-Coupled/genetics/metabolism
*Signal Transduction
Up-Regulation
Wnt Proteins/genetics/*metabolism