Bioinformatics Analysis And Molecular Docking Analysis Demonstrate ENMD-2076 As A Potent Therapeutic Drug Against Glioblastomas

Glioma is one of the most common primary malignant intracranial tumors in adults,accounting for 30% to 40% of brain tumors,and it is highly invasive.World Health Organization(WHO)classifies most of glioma as III~IV grade malignant tumors,and patients who have III~IV grade glioblastoma usually have limited prognosis.Although we have developed a series of comprehensive treatments which contained surgery,radiotherapy and chemotherapy,GBM patients’ prognosis is still unsatisfactory.In recent years,bioinformatics analysis enables us to understand the oncogenesis and development of gliomas from molecular level.Bioinformatics analysis helps researchers reveal therapeutic bio-targets in a systematic,accurate and effective way as well as illuminate theoretical basis of tumor occurrence and development,which makes it possible to study genetic changes and molecular mechanism of glioblastoma oncogenesis.With the help of bioinformatics,researchers could explore the hub driver genes underlying cellular mechanism of diseases and identify new diagnostic biomarkers or therapeutic targets from hub driver genes.Gene ontology(GO)analysis is a method to study biological process,molecular function and cell component of genes.Through GO annotation analysis,we can determine the biological functions,signaling pathways or cellular localization which have a close relationship with glioblastoma oncogenesis.Kyoto Encyclopedia of Genes and Genomes(KEGG)is a biological signaling pathway analysis,which can help us to find signal pathways that are significantly altered in glioblastoma samples.Protein-protein interaction network analysis(PPI)can help us systematically study cellular mechanism of glioblastoma as well as identify therapeutic targets.Gene set enrichment analysis(GSEA)was used to further explore altered signaling pathways of glioblastoma samples.Virtual drug screening and molecular docking have been widely used to identify inhibitors for targeted proteins,and a series of assessment modules are also used to evaluate pharmacological properties and affinity of drug candidates.ENMD-2076 is a type of oral bioavailable small molecular,cyclin-dependent kinase and aurora kinase inhibitor.It is also a fat-soluble,potent anti-tumor drug,which can easily pass through blood-brain barrier(BBB).In this study,we firstly identified hub driver genes of glioblastoma tissue by bioinformatics analysis,we then performed virtual screening and molecular docking to screen inhibitors for hub driver genes coding proteins.Subsequently,we performed a series experiments to evaluate therapeutic effect of the drug we identified,including MTT cell proliferation test,CFA colony formation test,in vitro scratch test,Transwell ventricular invasion test,western blot et al.Our main experimental results are as follows:(1)AURKA,NDC80,KIF4 A,NUSAP1,KDR are the hub genes accounting for glioblastomas oncogenesis.(2)Patients with low expression of AURKA,NDC80,KIF4 A,NUSAP1 gene have a better prognosis.(3)ENMD-2076 has a high binding affinity to Aurora kinase A and VEGFR-2,which is encoded by AURKA and KDR genes.ENMD-2076 is predicted to have no hepatotoxicity,good fat solubility,and easy to pass through blood-brain barrier(BBB).(4)ENMD-2076 inhibits proliferation and invasion of glioblastoma cells by inducing tumor cell apoptosis.(5)Animal experiments have proved that ENMD-2076 has an outstanding therapeutic effect regarding to glioblastoma.(6)ENMD-2076 inhibits invasion process of GBM cells by blocking the EMT process,inducing glioblastoma cell cycle arrest in G2-M phase,inducing glioblastoma cell apoptosis by reducing anti-apoptotic protein Bcl-2 and increasing pro-apoptotic proteins that cleave Caspase-3 and Bax.ENMD-2076 also inhibit proliferation,migration and apoptosis of GBM cells by inhibiting PI3K/AKT/m TOR signaling pathway.Conclusion:AURKA and KDR genes are key therapeutic targets for GBM.ENMD-2076 was found as a potent inhibitor for Aurora A kinase and VEGFR-2 through virtual screening technique.In vitro and in vivo studies demonstrated that ENMD-2076 is a promising,potent and safe drug in dealing with glioblastoma.