Expression of siRNA vector against HER2 in the human breast carcinoma SKBr-3 cells

AIM: To investigate the effect of HER2-targeted RNA interference (RNAi) on the growth of human breast carcinoma SKBr-3 cells. METHODS: HER2-targeted hairpin small interfering RNA (siRNA) genes were obtained by oligonucleotide synthesis and annealing of the complementary single strand DNAs. pSUPER vector harboring each of the above genes was transfected into SKBr-3 cells in vitro. The expression of HER2 gene in the transfected cells was examined by RT-PCR and indirect immunofluorescence assay, and the effect of the HER2-targeted siRNA on cell growth and proliferation was evaluated by cell counting and morphological observation. RESULTS: Both RT-PCR and indirect immunofluorescence assay revealed a remarkable decrease of HER2 expression in pSUPER-sihe1-and pSUPER-sihe2-transfected cells, but not in pSUPER vector transfected cells. Cell proliferation was dramatically inhibited after the expression of the HER2-targeted siRNAs as shown by microscopic observation and cell counting. CONCLUSION: HER2-targeted RNAi in SKBr-3 cells can effectively inhibit the expression of HER2 gene and the proliferation of the cells in vitro.