Identification And Characterization Of A Novel AMPK Activator With Potent Anti-glioma Efficiency

Background: Glioma is a primary intracranial neural tumor. Current chemotherapy treatment needs to overcome the blood-brain barrier and drug resistance. Despite progress in glioma clinical therapy options, such as surgery, radiation, chemotherapy, and combined modalities, the prognosis for malignant glioma patients remains dismal. Thus, it is necessary to develop novel and potent therapeutic agents for the treatment of glioma. Emerging evidence has revealed that AMP-activated protein kinase(AMPK) plays tumor suppressing roles in tumorigenesis.Methods:a) Cell growth was tested by MTT assay. Cell colony-forming assay was applied for detecting cell proliferation.b) The effect of Hh-009 on cell death was detected by Trypan Blue staining assay. We measured the cell apoptosis using Annexin V flow cytometry. Cell cycle distribution was explored by PI staining assay.c) The expression of signaling proteins(AMPK, ACC, AKT473, PDGFR and EGFR) was determined by Western blotting.d) U87 xenograft nude mice model was applied to test the potential anti-glioma activity by Hh-009 in vivo.Results:a) Hh-009 inhibited glioma cell growth and proliferation in a dose-dependent manner.b) Hh-009 efficiently induced glioma cell death and apoptosis. Hh-009 induced cell cycle arrest in glioma cells.c) Hh-009 activated AMPK signaling and induced autophagy activation in glioma cells. AMPK activation mediated Hh-009-induced AKT-m TOR in-activation and PDGFR/EGFR degradation in glioma cells.d) I.P. injection of Hh-009 significant inhibited U87 xenograft tumor growth in nude mice.Conclusions:Hh-009 potently inhibited glioma cell survival and growth, whiling inducing cell apoptosis, autophagy and cell cycle arrest. Hh-009 is a novel activator of AMPK, which causes degradation of PDGFR/EGFR and inhibition of m TORC1 signaling. Furthermore, Hh-009 administration suppressed the growth of solid tumor in U87 xenografts. Collectively, our findings suggest that targeting AMPK by Hh-009 could be a promising therapeutic approach for glioma prevention and therapy.