Inhibition of MAT 2A gene expression in hepatoma cell by short interfering RNAs

Objective To construct the expression vector pSlience-2.1-U6-siRNA that can express the short interfering RNAs (siRNA) against MAT2A gene and screen the effective target sequence of siRNA.Methods The four siRNA against MAT2A gene were transcript synthesized intracelluarly by expressed templates of plasmid vector pSilence-2.1-U6,and the target sequence of MAT2A gene was inserted into the upstream of the reporter gene in order to construct the recombinant plasmid vector plucF-MAT2A.The recombinant plasmid and siRNA -producing plasmid were cotransfected into 293 T cells by using Lipofectamine methods.The luciferase activity in cell lysate was quantitatively measured and the effective siRNA inhibiting the expression of luciferase was screened out.After the effective siRNA was transfected into Bel-7402 cells,the expression of MAT2A mRNA was semiquantitatively detected by semiquantitative RT-PCR.The MAT activity of hepatoma cells and apoptosis after transfection was measured.Results Two siRNA among the 4 synthesized siRNA displayed inhibitory effects on the lucifermase expression with the inhibitory rate being 81% and 89% respectively.The expression of MAT2A mRNA in Bel-7402 cells was specially ihhibited and the MAT activity decreased,and ultimately hepatic apoptosis induced.Conclusion The siRNA can inhibit the expression of MAT2A gene in hepatocellular carcinoma.