Effects and mechanism of adenovirus-mediated phosphatase and tension homologue deleted on chromosome ten gene on collagen deposition in rat liver fibrosis.
AIM
To evaluate the effects of phosphatase and tension homologue deleted on chromosome ten (PTEN) gene on collagen metabolism in hepatic fibrosis and the underlying mechanisms.


METHODS
Rat primary hepatic stellate cells (HSCs) and human LX-2 cells were transfected with adenovirus containing cDNA constructs encoding wild-type PTEN (Ad-PTEN), PTEN mutant G129E gene (Ad-G129E), and RNA interference constructs targeting the PTEN sequence PTEN short hairpin RNA to up-regulate and down-regulate the expression of PTEN. HSCs were assayed using fluorescent microscopy, real-time polymerase chain reaction, and western blotting. Moreover, a CCl4-induced rat hepatic fibrosis model was established to investigate the in vivo effects. Hematoxylin and eosin, and Masson's trichrome were used to assess the histological changes. The expression of collagen I and III was assessed using immunohistochemistry and western blot analysis.


RESULTS
Elevated expression of PTEN gene reduced serum levels of alanine transaminase and aspartate transaminase, decreased collagen deposition in the liver, and reduced hepatocyte necrosis. In contrast, knockdown of PTEN expression had an opposite effect, such as increased collagen deposition in the liver, and was molecularly characterized by the increased expression of matrix metalloproteinase (MMP)-13 (P < 0.01) and MMP-2 (P < 0.01), as well as decreased expression of the tissue inhibitor of metalloproteinase (TIMP)-1 (P < 0.01) and TIMP-2 (P < 0.01).


CONCLUSION
These data indicated that gene therapy using recombinant adenovirus encoding PTEN might be a novel way of treating hepatic fibrosis.
['Xie|Shu-Rui|SR|', 'An|Jun-Yan|JY|', 'Zheng|Li-Bo|LB|', 'Huo|Xiao-Xia|XX|', 'Guo|Jian|J|', 'Shih|David|D|', 'Zhang|Xiao-Lan|XL|']
D000256:Adenoviridae D000410:Alanine Transaminase D000818:Animals D001219:Aspartate Aminotransferases D002251:Carbon Tetrachloride D002460:Cell Line D024042:Collagen Type I D024061:Collagen Type III D015536:Down-Regulation D005109:Extracellular Matrix D055785:Gene Knockdown Techniques D015316:Genetic Therapy D005822:Genetic Vectors D055166:Hepatic Stellate Cells D006801:Humans D008103:Liver Cirrhosis D008297:Male D009154:Mutation D051059:PTEN Phosphohydrolase D061251:Primary Cell Culture D034741:RNA, Small Interfering D051381:Rats D014162:Transfection D015854:Up-Regulation
Phosphatase and tension homologue deleted on chromosome ten PTEN Collagen metabolism Hepatic fibrosis Hepatic stellate cells Gene therapy