Construction and expression of siRNA expression constructs targeting epidermal growth factor receptor
2005
Objective To construct the small interfering RNA(siRNA) expression constructs targeting epidermal growth factor receptor(EGFR) and express them in TJ905 human malignant cells. Methods Two target sequences from Receptor L domain and catalytic domain were selected to create two expression constructs using psiRNA-NeoG2. Furthermore, the siRNA constructs were transfected into TJ905 cells as mediated by Lipofectamin. Meanwhile, an antisense EGFR construct p-anti-hEGFR was set as control. Immunofluorescence and Western blot were performed to detect EGFR expression. Results With the successful construction of the two siRNA expression plasmids and the stable transfection to TJ905 cells, the expression of EGFR was down-regulated to 90% and 92% respectively, but to 82% in the anti-sense EGFR group. Conclusion The siRNA expression constructs targeting EGFR could specifically inhibit EGFR expression , and should be a new strategy in glioma gene therapy targeting EGFR.
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