Effect of lentivirus mediated HER2-shRNA on chemosensitivity of human ovarian cancer SKOV3 cells / 中国生化药物杂志

ABSTRACT

Objective To investigate the lentivirus-mediated RNA interference(RNAi) reduced human epidermal growth factor 2 (HER2) gene expression in ovarian cancer chemosensitivity.Methods Specifically targeted short hairpin RNA HER2 gene ( shRNA) lentiviral expression vector and negative control sequences lentiviral vector were constructed.stable expression of HER2-shRNA in SKOV3 cell line ( KD) and negative control cell line (NC) were established.The uninfected SKOV3cell lineserved as blank control(CON)group, sensitivity to changes in three kinds of cells to cisplatin chemotherapy in the HER2 gene expression was detected by RT-PCR and Western blot technique, CCK-8 assay CON and KD cells strains were established xenograft model in nude mice in each group to detect the expression of the HER2 protein, as well as by cisplatin chemotherapy tumor size. Results Expression results showed that a stable cell line HER2-shRNA and negative control cell lines had been successfully establish by fluorescence and flow cytometry.RT-PCR results showed KD cells relative HER2 mRNA expression levels compared with the NC group, the difference was statistically significant (P<0.01), whereas no significant difference when compared with the CON group, HER2 mRNA KD Group inhibition rate was (95.5 ± 0.12)%.After transduction specific HER2-shRNA ovarian cancer tumor cells, HER2 protein expression was significantly reduced, growth had slowed down significantly after cisplatin treatment ( P<0.01 ) .Conclusion Lentivirus-mediated HER2-shRNA can effectively inhibit the expression of the HER2 gene, increase chemosensitivity to cisplatin in ovarian cancer.