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Effects of c-myb antisense RNA on TGF-beta1 and beta1-I collagen expression in cultured hepatic stellate cells
Hui-Hui Ma1, Ji-Lu Yao, Gang Li
1Department of Infectious Diseases, the 3rd Affiliated Hospital, Sun-Yat Sen University, Guangzhou 510630, Guangdong Province, China. lucam@.com
Aim:
To investigate the effects of c-myb antisense RNA on cell proliferation and the expression of c-myb, TGF-beta1 and beta1-I collagen in cultured hepatic stellate cells (HSC) from rats.
Methods:
Recombinant retroviral vector of c-myb antisense gene (pDOR-myb) was constructed, and then transfected into retroviral package cell line PA317 by means of DOTAP. The pseudoviruses produced from the resistant PA317 cells were selected with G418 to infect HSCs isolated from rat livers. The cell proliferation was measured by 3-[4, 5-Dimethylthiazolzyl]-2, 5-diphenyl tetrazo-dium bromide(MTT) method. The expression of c-myb, alpha (1)-I collagen and TGF-beta1 mRNA, and c-myb protein in HSCs was detected with semi-quantitive reverse transeription-polymerase chain reaction (RT-PCR) and Western-blot respectively.
Results:
HSCs from rats were isolated successfully with the viability >98%. In the pDOR-myb infected HSCs, the c-myb protein expression, cell proliferation,and alpha (1)-I collagen and TGF-beta1 mRNA expression were repressed significantly compared with their corresponding control groups (P<0.01).
Conclusion:
c-myb plays a key role in activation and proliferation of HSC. c-myb antisense RNA can inhibit cell proliferation, alpha (1)-I collagen and TGF-beta1 mRNA expression, suggesting that inhibition of c-myb gene expression might be a potential way for the treatment of liver fibrosis.
