The Effects And Mechanisms Of Dihydroartemisinin On Apoptosis And Migration Of Human Malignant Glioma Cells

Background: Malignant glioma is a highly lethal malignant brain tumor that accounts for 80% of adult primary brain tumors.Despite aggressive surgical resection followed by radiation and chemotherapy,the median survival time of patients with glioblastoma multiforme(World Health Organization grade IV)is only 12–14 months.Its aggressiveness and critical health care matter mainly arise from the extensive proliferation and insidious invasion capabilities of malignant glioma cells.To date,the search for novel and more efficient chemo-agents against this deadly disease has become a major research focus in the field of glioblastoma multiforme treatment.Dihydroartemisinin(DHA)is one of the main active metabolites of arteminisin.In recent years,more and more studies have revealed that DHA has significant anti-cancer effects,especially for ovarian cancer.In glioma cells,previous studies have reported that DHA has been shown to inhibit proliferation,metastasis,and invasion,and enhance radio sensitivity,but these molecular mechanisms need to be further investigated.Aims: To explore the effects and potential mechanisms of DHA on malignant glioma cells,searching for new potential drug to treat the proliferation,migration and invasion of glioblastoma in clinical treatment and providing some basic study for further studies in animal experiments and pre-clinical studies.Methods: The human normal glial cell lines(HEB)and glioma cell lines(U87,U251,U343 and HS683)were treated with DHA.MTT assay was used for detecting the effect of DHA on glioma cells viability.The cells morphological changes were observed by microscope.Hoechst 33258 Staining and flow cytometry were used for determining the effect of DHA on the apoptosis of glioma cells.Colony formation was used for studying the long-term effect of DHA on glioma cells growth,cell scratch and transwell assay were used for evaluating the migration and invasion abilities of malignant glioma cells in vitro;Q-PCR for detecting the relative quantity of miR-21 and RECK;miR-21 transfection technology was used to assay its role in DHA killing glioma cells;Western-blot was used to value the changes of key proteins associated with AKT-p53 and EMT pathways.The relationship between RECK expression and clinicopathological features of brain glioma patients is analyzed with SPSS software.Results: Dihydroartemisinin(DHA)could inhibit the growth and viability in malignant glioma cells.After Dihydroartemisinin(DHA)treatment,the results showed that DHA could significantly induce malignant glioma cells(U251,U87,U343 and HS683)apoptosis through AKT-p53 apoptosis pathway and cells death through miR-21 down-regulation.Correspondingly,Dihydroartemisinin(DHA)could suppress glioma cells migration and invasion through regulating EMT process associated proteins,down-regulating MMP-2 and N-Cadherin,up-regulating RECK protein,and the expression of RECK has a close correlation with the grades of malignant glioma.Conclusion: Our findings demonstrated that AKT-p53 pathway,miR-21 and RECK play important roles in Dihydroartemisinin(DHA)inducing malignant glioma cells apoptosis,death and inhibiting cells migration,revealing that DHA could be developed as a potential clinical therapeutic drug,with a view to improving the poor treatment outcomes of glioblastoma.