EFFECT OF siRNA SILENCING EGFR GENE ON INVASIVE POWER OF OVARIAN CANCER CELLS
Huaihua Song
Abstract
Huaihua Song
Abstract
Objective To investigate the effect of RNA interference(RNAi) on the invasion power of ovarian cancer cell line SKOV3 in vitro through the gene expression of small interference silence epidermal growth factor receptor(EGFR). MethodsThe siRNA plasmid of target EGFR gene and negative-control plasmid were constructed in vitro,which were transfected to SKOV3 cells mediated by Lipofectamin 2000.The study consisted of three groups: blank control;non-specific siRNA transfected(NS-siRNAT) and specific siRNA transfected(S-siRNAT).The expressions of EGFR mRNA and protein were detected by reverse transcription polymerase chain reaction(RT-PCR) and immunocytochemical tecnique.The proliferation of SKOV3 cells was detected by clone formation assay and in vitro invasive power of the cells detected by Transwell assay. Results The expression in SsiRNAT group was markedly weaken than that in the orther two groups in terms of EGFR mRNA(F=50.45;q=12.87,11.72;P0.01),the protein(F=61.54;q=14.58,11.46;P0.01),the proliferation of the cells(F=130.12;q=21.12,18.15;P0.01) and in vitro invasive power(F=34.09;q=11.26,8.34;P0.01). Conclusion siRNA can effectively suppress the expression of EGFR gene and lead to the suppression of proliferation and invasiveness of SKOV3 cells.
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Objective To investigate the effect of RNA interference(RNAi) on the invasion power of ovarian cancer cell line SKOV3 in vitro through the gene expression of small interference silence epidermal growth factor receptor(EGFR). MethodsThe siRNA plasmid of target EGFR gene and negative-control plasmid were constructed in vitro,which were transfected to SKOV3 cells mediated by Lipofectamin 2000.The study consisted of three groups: blank control;non-specific siRNA transfected(NS-siRNAT) and specific siRNA transfected(S-siRNAT).The expressions of EGFR mRNA and protein were detected by reverse transcription polymerase chain reaction(RT-PCR) and immunocytochemical tecnique.The proliferation of SKOV3 cells was detected by clone formation assay and in vitro invasive power of the cells detected by Transwell assay. Results The expression in SsiRNAT group was markedly weaken than that in the orther two groups in terms of EGFR mRNA(F=50.45;q=12.87,11.72;P0.01),the protein(F=61.54;q=14.58,11.46;P0.01),the proliferation of the cells(F=130.12;q=21.12,18.15;P0.01) and in vitro invasive power(F=34.09;q=11.26,8.34;P0.01). Conclusion siRNA can effectively suppress the expression of EGFR gene and lead to the suppression of proliferation and invasiveness of SKOV3 cells.
Key concepts: Transfection, RNA interference, Molecular biology, Gene silencing, Small interfering RNA, In vitro, Biology, Gentamicin protection assay