Effects of c-myb antisense RNA on TGF-β1 and α1-I collagen expression in cultured hepatic stellate cells

AIM:To investigate the effects of c-myb antisense RNA oncell proliferation and the expression of c-myb,TGF-131 andα1-I collagen in cultured hepatic stellate cells (HSC) from rats.METHODS:Recombinant retroviral vector of c-myb antisensegene (pDOR-myb) was constructed,and then transfectedinto retroviral package cell line PA317 by means of DOTARThe pseudoviruses produced from the resistant PA317 cellswere selected with G418 to infect HSCs isolated from ratlivers.The cell proliferation was measured by 3-[4,5-Dimethylthiazolzyl]-2,5-diphenyl tetrazo-dium bromide(MIT) method.The expression of c-myb,α1-I collagen andTGF-β1 mRNA,and c-myb protein in HSCs was detectedwith semi-quantitive reverse transeription-polymerase chainreaction (RT-PCR) and Western-blot respectively.RESULTS:HSCs from rats were isolated successfully withthe viability>98%.In the pDOR-myb infected HSCs,the c-myb protein expression,cell proliferation,and α1-I collagenand TGF-β1 mRNA expression were repressed significantlycompared with their corresponding control groups (P<0.01).CONCLUSION:c-myb plays a key role in activation andproliferation of HSC.c-myb antisense RNA can inhibit cellproliferation,α1-I collagen and TGF-β1 mRNA expression,suggesting that inhibition of c-myb gene expression mightbe a potential way for the treatment of liver fibrosis.