Transcriptional profiling and Wnt signaling activation in proliferation of human hepatic stellate cells induced by PDGF-BB.
OBJECTIVE
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/calmodulindependent 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/Ca(2+) signaling pathway is active in hTERT HSCs and may participate in HSC activation and proliferation.
Shin
Hye Won
HW
Department of Pathology, Seoul National University College of Medicine, Korea.
Park
Soo Young
SY
Lee
Kyoung Bun
KB
Shin
Eun
E
Nam
Suk Woo
SW
Lee
Jung Young
JY
Jang
Ja June
JJ
D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D000077214:Becaplermin; D015153:Blotting, Western; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D051157:Frizzled Receptors / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D055166:Hepatic Stellate Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D020574:Proto-Oncogene Proteins c-sis; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D015854:Up-Regulation; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism*
D043925:Angiogenesis Inducing Agents; C121168:FZD10 protein, human; D051157:Frizzled Receptors; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D043562:Receptors, G-Protein-Coupled; D051153:Wnt Proteins; D000077214:Becaplermin; C517657:CAMK2A protein, human; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2