Specific siRNA targeting EGFR enhances ovarian cancer cell line SKOV-3 apoptosis
Hongling Zhang, Aiping Chen, Hui Song, Ruirui Yang
Abstract
Hongling Zhang, Aiping Chen, Hui Song, Ruirui Yang
Abstract
Objective To observe the influence of specific short hairpin siRNA targeting EGFR gene on apoptosis of human ovarian cancer Skov-3 cells in vitro. Methods A plasmid of a short hairpin siRNA targeting EGFR was constructed, and it was transfeeted into Skov-3 cell line by lipofectamine 2000. Human ovarian carcinoma cells of the line Skov-3 were cultured and divided into 3 groups: control group; non-specific group, transfected with non-specific plasmid vector; and specific group, transfected with specific small hairpin RNA expression vector. The expression of EGFR mRNA and protein were examined by RT-PCR and immunocytochemistry, Flow cytometry (FCM) was adopted to analyze quantitatively apoptotic cells in each group. Results After transfection of pshRNA-EGFR, mRNA and protein levels of EGFR gene in Skov-3 cells were obviously reduced. Flow cytometry analysis revealed that apoptosis could be induced in Skov-3 cells line transfected with pshRNA-EGFR in a time-dependent manner, no obvious apoptosis were detected in control group and non-specific group. Conclusion The plasmid expressive vector target at EGFR in our study is capable of suppressing EGFR expression of human ovarian cancer Skov-3 cells and inducing apoptosis, which provide a new way for the gene therapy of human ovarian cancer. Key words: Genes,erbB-1; Ovarian neoplasms; RNA interference; Apoptosis
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Objective To observe the influence of specific short hairpin siRNA targeting EGFR gene on apoptosis of human ovarian cancer Skov-3 cells in vitro. Methods A plasmid of a short hairpin siRNA targeting EGFR was constructed, and it was transfeeted into Skov-3 cell line by lipofectamine 2000. Human ovarian carcinoma cells of the line Skov-3 were cultured and divided into 3 groups: control group; non-specific group, transfected with non-specific plasmid vector; and specific group, transfected with specific small hairpin RNA expression vector. The expression of EGFR mRNA and protein were examined by RT-PCR and immunocytochemistry, Flow cytometry (FCM) was adopted to analyze quantitatively apoptotic cells in each group. Results After transfection of pshRNA-EGFR, mRNA and protein levels of EGFR gene in Skov-3 cells were obviously reduced. Flow cytometry analysis revealed that apoptosis could be induced in Skov-3 cells line transfected with pshRNA-EGFR in a time-dependent manner, no obvious apoptosis were detected in control group and non-specific group. Conclusion The plasmid expressive vector target at EGFR in our study is capable of suppressing EGFR expression of human ovarian cancer Skov-3 cells and inducing apoptosis, which provide a new way for the gene therapy of human ovarian cancer. Key words: Genes,erbB-1; Ovarian neoplasms; RNA interference; Apoptosis
Key concepts: Transfection, Lipofectamine, Small hairpin RNA, Apoptosis, Flow cytometry, Molecular biology, Cancer research, RNA interference