Effects Of HER-2/neu SiRNA-mediated Gene Silencing On Cell Proliferative Activity In Human Meningioams

Objective:In this study, we established primary cultures from human meningioma tissues.Investigating the effect on cell proliferative activity after transient transfection withspecific HER-2/neu small interfering RNA(HER-2siRNA) silencing in HER-2(Human epidermal growth factor receptor-2) positive cells.Methods:The subject collected surgical resection of fresh human meningioma tissue werecultured and screened by immunocytochemistry HER-2antibody positive cases assubjects,transient transfection of HER-2positive meningioma cells with specificHER-2siRNA,the experiment was divided into four groups: specific Her-2siRNAgroup; negative control siRNA group; liposome group; blank control group. Afterconfirming the successful transfection, MTT assay was employed to evaluate cellviability. Ki-67protein levels which indicate cell proliferation activity was analyzedby immunocytochemistry.While the expression of HER-2gene was evaluated bysemi-quantitative RT-PCR, immunoblot analysis and immunocytochemistry.Explore meningioma targeting blockage of the HER-2gene expression changes atthe cellular level, RNA and protein levels, to observe the relationship between theHER-2gene and meningioma biological behavior.Results:MTT assay revealed that silencing HER-2gene with the specificity of HER-2siRNA suppressed primary cultured human meningioma cellular proliferation from48h after transfection.72h after transfection the strongest inhibition of cellproliferation inhibition rate of50.08%, Inhibition rate in96h after transfection and72h after transfection,there was no significant difference;72h after transfection,RT-PCR results showed that HER-2mRNA relative expression levels of HER-2siRNA group compared to before transfection significantly reduced; after transfection72h,immunocytochemistry results showed that the HER-2protein and Ki-67protein expression in specific HER-2siRNA group meningioma cells significantly reducedcompared with before transfectionï¼›after transfection72h,western blot results showedthat the specificity of HER-2siRNA group meningioma cells relative expressionlevels of HER-2protein than before transfection significantly reduced.Conclusion:At the cellular level, the specificity of HER-2siRNA can effectively inhibit the invitro cultured human meningioma cell proliferation, RNA and protein levels may alsoeffectively inhibit the expression of HER-2gene. HER-2gene was closely related tothe biological behavior of primary cultured human meningioma cells.This studysuggests the possibility of refractory or recurrent meningioma therapeutics using thespecificity HER-2siRNA.