[Construction and expression of siRNA expression constructs targeting epidermal growth factor receptor].
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.


CONCLUSIONS
The siRNA expression constructs targeting EGFR could specifically inhibit EGFR expression, and should be a new strategy in glioma gene therapy targeting EGFR.
Kang
Chunsheng
C
Department of Neurosurgery, Tianjin Medical University, General Hospital and Laboratory of Neuro-oncology, Tianjin Neurological Institute, Tianjin, 300052 P. R. China.
Pu
Peiyu
P
Wang
Guangxiu
G
Dong
Lun
L
Li
Yanhe
Y
Wang
Hu
H
D015153:Blotting, Western; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection / Q000379:methods
D034741:RNA, Small Interfering; D066246:ErbB Receptors