An experiment study about the apoptosis effect induced by adenovirus mediated HIV1-Vpr on human glioma cells in vitro

Objective To study the effect of recombinant adenovirus mediated transfection of human immunodeficiency virus type 1 viral protein R (HIV1 - Vpr) on apoptosis of glioma U251 cells in vitro.Methods U251 cells under culture were divided into the normal control group, mock group and experiment group.After 24 hours of culture, the mock and experiment group were transfected with mock vector(rAd5 -null)and the recombinant adenovirus carrying HIV1 -Vpr (rAd5 - Vpr) respectively at a multiplicity of infection (MOI) of 50.The proliferation activity of cells was detected by MTT assay, the apoptosis effect and the change of cell cycle were determined by Hoechst stain and flow cytometry (FCM) technology respectively. The expression of related proteins was revealed by the method of Western blot. Results rAd5 - Vpr could inhibit the proliferation of U251 cells, and this effect may start48 hours after transfection and increase with the prolonged time (P 0.05); The results of Western blot showed that after 72 hours of transfection the Vpr protein expression was observed, meanwhile, the Bcl-2 protein showed decreased expression while in the Bax,Caspase3 and Fas - L proteins increased expression Conclusion rAd5 - Vpr could inhibit the proliferation of U251cells and induce cell cycle G2 arrest and apoptosis in vitro. Key words: Glioma; Apoptosis; Human immunodeficiency virus type 1; Viral protein regulatory; Recombinant adenovirus