Association of differentially expressed genes with activation of mouse hepatic stellate cells by high-density cDNA microarray.
OBJECTIVE
To characterize the gene expression profiles associated with activation of mouse hepatic stellate cell (HSC) and provide novel insights into the pathogenesis of hepatic fibrosis.


METHODS
Mice HSCs were isolated from BALB/c mice by in situ perfusion of collagenase and pronase and single-step density Nycodenz gradient. Total RNA and mRNA of quiescent HSC and culture-activated HSC were extracted, quantified and reversely transcribed into cDNA. cDNAs from activated HSC were labeled with Cy5 and cDNAs from the quiescent HSC were labeled with Cy3, which were mixed with equal quantity, then hybridized with cDNA chips containing 4000 genes. Chips were washed, scanned and analyzed. Increased expression of 4 genes and decreased expression of one gene in activated HSC were confirmed by reverse transcription-polymerase chain reaction (RT-PCR).


RESULTS
A total of 835 differentially expressed genes were identified by cDNA chip between activated and quiescent HSC, and 465 genes were highly expressed in activated HSC. The differentially expressed genes included those involved in protein synthesis, cell-cycle regulation, apoptosis, and DNA damage response.


CONCLUSIONS
Many genes implicated in intrahepatic inflammation, fibrosis and proliferation were up-regulated in activated HSC. cDNA microarray is an effective technique in screening for differentially expressed genes between two different situations of the HSC. Further analysis of the obtained genes will help understand the molecular mechanism of activation of HSC and hepatic fibrosis.
Liu
Xiao-Jing
XJ
Laboratory of Department of Internal Medicine, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China. xiaojingliu67@yahoo.com
Yang
Li
L
Luo
Feng-Ming
FM
Wu
Hong-Bin
HB
Qiang
Qu
Q
D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology*; D008103:Liver Cirrhosis / Q000503:physiopathology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction