Disruption of connective tissue growth factor by short hairpin RNA inhibits collagen synthesis and extracellular matrix secretion in hepatic stellate cells.
OBJECTIVE
Connective tissue growth factor (CTGF) plays a key role in the pathogenesis of liver fibrosis. This study aimed at investigating that the disruption of CTGF expression by short hairpin RNA (shRNA) could modulate the production of fibrosis-related components in hepatic stellate HSC-T6 cells.


METHODS
Three plasmids expressing individual shRNA were constructed and transfected into HSC-T6 cells respectively. The levels of CTGF and transforming growth factor beta1 (TGF-beta1) expression were determined by reverse transcriptase-polymerase chain reaction and Western blot assays. Furthermore, the production of collagens and extracellular matrix (ECM) proteins, including type III procollagen (PC III), type IV collagen (collagen IV), laminin (LN) and hyaluronic acid (HA) in the CTGF-disrupted cells, were analysed by radioimmunoassay.


RESULTS
Following transfection with the pEGFP-CTGFshRNA1, the expression of CTGF was disrupted in HSC-T6 cells. While the knockdown of CTGF expression failed to modulate the expression of TGF-beta1, it did significantly reduce the production of PC III, collagen IV, LN and HA by HSC-T6 cells.


CONCLUSIONS
Our data suggest that shRNA-mediated disruption of CTGF expression can attenuate ECM synthesis. Potentially, our findings may aid in the design of a CTGF-based new therapy for treatment of hepatic fibrosis.
Yuhua
Zhu
Z
Department of Gastroenterology, Provincial Hospital affiliated to Shandong University, Shandong, China.
Wanhua
Ren
R
Chenggang
Sun
S
Jun
Shi
S
Yanjun
Wang
W
Chunqing
Zhang
Z
D000818:Animals; D002460:Cell Line; D003094:Collagen / Q000096:biosynthesis*; D055513:Connective Tissue Growth Factor; D015536:Down-Regulation; D005109:Extracellular Matrix / Q000378:metabolism; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D017874:Immediate-Early Proteins / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D007797:Laminin / Q000096:biosynthesis*; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids; D034622:RNA Interference; D051381:Rats; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*
C526316:CCN2 protein, rat; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D007797:Laminin; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor; D006820:Hyaluronic Acid; D003094:Collagen