Effect of Adenovirus Vector-mediated RNA Interference on Expression of Hepatocyte Growth Factor Receptor c-Met in Glioma U251 Cells

Objective To investigate the effects of Adenovurus vecror-mediated RNA intereference (RNAi) on the expression of hepatocyte growth factor (HGF) receptor, c-met in glioma U251 cells and apoptosis of glioma U251 cells. Methods Human U6 promotor and the HU6shmet, fragment with c-Met reverse complement target sequence were obtained by PCR. The Hu6shmet was packaged in adenovirus and transduced into U251 cells. The expressions of c-Met mRNA and protein were determined respectively by RT-PCR and Western blot. The apoptosis of Glioma U251 cells was determined by flow cytometry. Results The Recombined adenovirus vector with human U6 promotor and c-Met reverse complement target sequence (named rAdUshmet 1 and rAdUshmet 2) were obtained. The c-Met mRNA and protein expression levels were significantly lower in the U251 cells transduced with the rAdUshmet 1 and rAdUshmet 2 than those in U251 cells without transduction with adenovirus, U251 cells transduced with recombined adenovirus with U6 small inteterence control(rAdUsicon) and U251 cells transduced with recombined adenovirus with green fluorescent protein(rAdGFP) (P0.01). The apoptosis rates of the cells were (13.5±4.2)% and (28.2±5.6)% respectively in U251 cells transduced with rAdUshmet 1 and rAdUshmet 2, and were significantly higher than those in U251 cells without transduction with adenovirus, U251 cells transduced with rAdUsicon or with rAdGFP (P0.05). Conclusions The rAdUshmet can inhibit c-Met expression and induce apoptosis of glioma cells, block the HGF-c-Met signal pathway partially and may act as an effective vector for gene therapy of glioma.