The Inhibitory Effect Of Silencing Her-2by ShRNA On The Expressions Of VEGF-A、IL-8in Osteosarcoma Cell Line

Objective To study the regulatory effect of her-2gene by shRNA onthe expression of VEGF-A、IL-8in osteosarcoma cell line saos-2.Methods Her-2recombinant expression vector was constructed andtransfected into osteosarcoma cell line saos-2by liposome-mediatedtransfection; the well-transfected saos-2cells were screened out usingG418. Fluorescence microscopy and flow cytometry were used to observethe transfection efficiency. The expression levels of her-2mRNAs andprotein were detected by RT-PCR and Western-blot, while the expressionlevels and the secretion levels of VEGF-A, IL-8by Western-blot andELISA.Results Her-2shRNA expression plasmid was successfullyconctructed and verified by sequencing. Her-2shRNA expression plasmidwas stably transfected into osteosarcoma cell line saos-2and inhibited theexpression of her-2gene, of which the inhibiting rates to her-2mRNAexpression and protein expression were63.05%and62.59%. RT-PCR andELISA assay, compared with blank control group and negative control group, VEGF-A, IL-8mRNA and secretion levels were significantlyinhibited in the her-2shRNA-transfected group (P<0.01).Conclusion Inhibition of her-2gene expression can decrease theexpression levels of VEGF-A, IL-8significantly, implying that her-2might involve in the regulation of VEGF-A, IL-8in osteosarcoma cells andher-2gene might be used as a potential target for exploring the molecularof osteosarcoma angiogenesis.