SiRNA Targeting Human IGF-â…¡ Gene Driven By Recombinant RNA Polymerase â…¡ HAFP/hTERT Dual Promoters Specifically Suppresses The Growth Of Hepatocellular Carcinoma Cells

Objective: To observe the effect of siRNA expression vector driven by recombinant RNApolymerase â…¡ hAFP/hTERT dual promoters on the growth of hepatocellular carcinoma cells.Methods: The siRNA expression vectors driven by the hAFP and hTERT promoters weretransfected into Huh-7cells and L-02cells and their IGF-â…¡ mRNA expression levels weredetected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) at48h aftertransfection. The proliferations were evaluated by using MTS and the changes in cell cycle andapoptosis were assessed by using flow cytometry. The protein levels of IGF-â…¡, PCNA, cyclinE2, cyclin D2, cdc2and Bcl-2were annalsyzed by western blot.Results:â‘ The expression of IGF-â…¡ mRNA in Huh-7cells was reduced by86%approximately; the proliferations of Huh-7cells were remarkablely inhibited; their number inG1phase significantly increased, but S phase remarkablely decreased and the percentage ofearly apoptotic cells, late apoptotic cells and total apoptotic cells(early apoptotic cells plus lateapoptotic cells) in Huh-7cells transfected with siRNA were much higher than that in Huh-7cells transfected with blank control or negative control group, respectively; the protein levels ofIGF-â…¡, PCNA, cyclin E2, cyclin D2, cdc2and Bcl-2in Huh-7cells significantly decreased.â‘¡There were no differences in the above parameters with respect to L-02cells.Conclusion: The siRNA expression vector driven by recombinant RNA polymerase â…¡hAFP/hTERT dual promoters can specifically suppress the IGF-â…¡ mRNA expression andgrowth of hepatocellular carcinoma cells and it may be involved in decreasing the protein levelsof IGF-â…¡, PCNA, cyclin E2, cyclin D2, cdc2and Bcl-2through down-regulation of IGF-â…¡expression, and it could become a new approach for gene therapy of hepatocellular carcinoma.