EFFECT OF SILENCING EGFR GENE EXPRESSION BY RNA INTERFERENCE ON THE CHEMOSENSITIVITY OF OVARIAN CANCER CELL LINE skov3

Objective To construct expression vector of EGFR specific siRNA and transfect it into human ovarian cancer cell line skov3 and then investigate the effect of RNA interference(RNAi) on the target gene expression and chemosensitivity of the cell line. Methods EGFR shRNA was synthesized in vitro and then transfected into skov3 cell to set up the EGFR shRNA transfectant cell line.The normal control cell line and non-specific transfectant cell line were also included in the study.RT-PCR and immunocytochemistry were performed to detect the inhibitive effect of RNAi on mRNA level and protein level,respectively.The chemosensitivity to cisplatin was detected by MTT method. Results The expression of EGFR mRNA of sequence specific shRNA targeting EGFR was obviously down-regulated(F=87.73;q=16.81,15.33;P0.01) and the same happened to the protein expression(F=69.29;q=14.71,13.57;P0.01),while the chemosensitivity to cisplatin was increased by three times. Conclusion The EGFR specific shRNA expression vector can effectively suppress the expression level of EGFR gene of skov3 cells,and increases its chemosensitivity to cisplatin.