Efficacy of genetically engineered oncolytic virus on glioma stem cells in vitro

Abstract

Objective Exploring the effect of endostatin and angiostatin(Endo-Angio) fusion genemodified herpes simplex virus ( VAE ) on glioma stem cells ( GSCs ) in vitro. Method Surgical specimens of human high grade glioma ( WHO Ⅲ, Ⅵ) were collected, and GSCs were isolated under the culture conditionsoriginally designed for selective expansion of neural stem cells. In order to identify GSCs,the number of CD133 positive cells in GSCs were detected by flow cytometry; self - renewal capacity and the expression of Nestin, GFAP, NF, and MAG of GSCs were detected by monoclonal forming and immunofluorescence assay respectively. After that, GSCs viability was detected by MTS assay. Expression of Endo - Angio fusion gene at mRNA and protein level was detected by RT - PCR and Western blot. At last,the efficacy of fusion protein to human brain microvascular endothelial cells (HBMEC) and the biological properties of GSCs were detect after infected by VAE. Results ( 1 )4 GSCs isolated from 20 surgical specimens could suspend growth and had the ability of self-renewal and multipotential differentiation,and could express neural stem cells marker,CD133 and Nestin. (2)VAE could infect GSCs and significantly inhibit the viability of GSCs( P ＜0.05). (3) Significant expression of Endo-Angio fusion gene was observed and it could inhibit HBMEC proliferation( P ＜0.05). (4) Residual viable cells lost the ability of self - renewal and adherent differentiation. Conclusions In vitro, VAE can inhibit the activity of GSCs and the expression of exogenous Endo - Angio fusion gene can inhibit HBMEC proliferation. VAE can be used as a novd virus -gene therapy strategy for glioma. Key words: Glioma; Neoplastic stem cells; Endostatins; Angiogenesis inhibitors; Virus - gene therapy