Quantified analysis of small interfering RNA silences transient expression of reporter gene in human hepatocarcinomal cells

Aim To investigate the feasibility and efficiency of small interfering RNA(siRNA) inhibiting transient gene expression in human hepatocarcinomal cells.Methods The siRNA degradation after incubation in vitro was observed by gel electrophoresis.The cellular kinetics of transfecting siRNA was screened with Cy3 labeling.The HepG2 cells were transiently co-transfected of green fluorescent protein(GFP) geneand synthesized siRNA.The quantified reporter gene(GFP) expression suppressed by specific siRNA was assayed by flow cytometery,Western blot and real-time PCR.Results The siRNA sustained relative stability for a long time in culture medium and transfected cells.Compared with mock and non-specific siRNA,GFP siRNA dominantly suppressed the expression of GFP protein and mRNA(P0.05).Conclusion The specific siRNA can high-efficiently silence the transient expression of transfectd gene in human hepatocarcinomal cells,and provides a new strategy for its gene therapy.