Serum GFAP and IGFBP-2 levels on mutated AKT/KRAS induced glioma in the mouse

Abstract

IntroductionGlioblastomamultiforme is a highly malignant brain tumor which is categorized as invasive tumor due to poor prognosis and difficult to diagnose at earlier stage of the tumor. The development of an animal model for glioma using RCAS/TVA system is a powerful tool to address on gene function and therapy, tumorigenesis and oncogenesis. ObjectiveTo develop an animal model for glioma using RCAS/TVA system and measure two proteins which are glial fibrillary artificial protein (GFAP) and insulin growth factor binding protein 2 (IGFBP2) as markers of the progression of glioma formation. MethodsPlasmid DNA carrying oncogenes of glioma (AKT and KRAS) was transformed into competent E.coli and the successful transformed plasmid DNA was extracted. The purity and concentration of plasmid DNA was obtained and cut by restriction enzyme (Hind III and Sma I). Then, plasmid DNA was transfected into chicken fibroblast DF-1 cell. The transfected DF-1 cell was confirmed by ALV p27 ELISA and western blot. Newborn mice carrying Nestin/T-va strains at the age of 24 hours were injected with infected DF-1 cell and latency period to develop glioma was within 3-5 weeks after injection. Blood was obtained from the tail at the age of 4weeks until week 14. Serum GFAP and IGFBP2 were measured by ELISA. ResultGlioma was successfully developed in the mouse model using a mutated AKT/KRAS plasmid DNA. Serum GFAP and IGFBP2 were significantly higher in mice injected with infected DF-1 cell carrying oncogenes for glioma (p<0.05)compared to mice without injection. ConclusionGFAP and IGFBP-2 have the potential as serum biomarkers for glioma in animal model. Thus, this reliable blood-based tumor biomarkers could significantly improve clinical diagnosis and research studies in glioma patients.