Saydam O.Glauser, D.L.Heid I.Turkeri G.Hilbe, M.Jacobs, A.H.Ackermann, M.Fraefel, C.2016-02-082016-02-08200515250016http://hdl.handle.net/11693/23963In primary glioblastomas and other tumor types, the epidermal growth factor receptor (EGFR) is frequently observed with alterations, such as amplification, structural rearrangements, or overexpression of the gene, suggesting an important role in glial tumorigenesis and progression. In this study, we investigated whether posttranscriptional gene silencing by vector-mediated RNAi to inhibit EGFR expression can reduce the growth of cultured human gli36 glioma cells. To "knock down" EGFR expression, we have created HSV-1-based amplicons that contain the RNA polymerase III-dependent H1 promoter to express double-stranded hairpin RNA directed against EGFR at two different locations (pHSVsiEGFR I and pHSVsiEGFR II). We demonstrate that both pHSVsiEGFR I and pHSVsiEGFR II mediated knock-down of transiently transfected full-length EGFR or endogenous EGFR in a dose-dependent manner. The knock-down of EGFR resulted in the growth inhibition of human glioblastoma (gli36-luc) cells both in culture and in athymic mice in vivo. Cell cycle analysis and annexin V staining revealed that siRNA-mediated suppression of EGFR induced apoptosis. Overall HSV-1 amplicons can mediate efficient and specific posttranscriptional gene silencing. Copyright © The American Society of Gene Therapy.EnglishEGFRHSV-1 ampliconsPosttranscriptional gene silencingTumor growthDNA directed RNA polymerase IIIdouble stranded RNAepidermal growth factor receptorlipocortin 5short hairpin RNAsmall interfering RNAvirus vectorampliconanimal experimentanimal modelantineoplastic activityapoptosisarticlecancer inhibitioncell cyclecontrolled studygene functiongene targetinggenetic transfectiongliomaHerpes simplex virus 1humanhuman cellin vivo studynonhumannude mouseposttranscriptional gene silencingpromoter regionprotein domainprotein expressionRNA interferencetumor cell cultureviral gene delivery systemGene Expression Regulation, NeoplasticGene TherapyGene Transfer TechniquesGenes, erbB-1Genetic VectorsGlioblastomaHerpesvirus 1, HumanHumansReceptor, Epidermal Growth FactorRNA InterferenceRNA, Small InterferingTumor Cells, CulturedAnimaliaHuman herpesvirus 1Mus musculusArticle10.1016/j.ymthe.2005.07.534