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Published on: January 4, 2012
[Construction and expression of siRNA expression constructs targeting epidermal growth factor receptor]
Chunsheng Kang1, Peiyu Pu, Guangxiu Wang
1Department of Neurosurgery, Tianjin Medical University, General Hospital and Laboratory of Neuro-oncology, Tianjin Neurological Institute, Tianjin, 300052 P. R. China.
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.
