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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,α_1-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 α_1-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,α_1-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.
AIM: To investigate the effects of c-myb antisense RNA once once proliferation and the expression of c-myb, TGF-131 and al-I collagen in cultured hepatic stellate cells (HSC) from rats. METHODS: Recombinant retroviral vector of c-myb antisensegene (pDOR-myb) was constructed, and then transfected with retroviral package cell line PA317 by means of DOTARThe pseudoviruses produced from the resistant PA317 cells with 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, α 1 -I collagen and TGF- β 1 mRNA, and c-myb protein in HSCs was detected with semi-quantitive reverse transection-polymerase chain reaction Mycoprotein, cell proliferation, and α_1-I collagen and TGF-β1 were detected by RT-PCR and Western-blot respectively.RESULTS: HSCs from rats were isolated successfully with the viability> 98% β1 mRNA expression were repressed significan suggesting that inhibition of tc-myb antisense RNA can inhibit cell proliferation, 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.