Silencing DRP1 Inhibits Glioma Cells Proliferation And Invasion Via RHOA/ROCK1 Pathway

Objectives: Dynamin-related protein 1(Drp1)is a newly discovered therapeutic target for tumor initiation,migration,proliferation,and chemosensitivity,while its molecular mechanism in glioma is unclear.This study aimed to determine the expression of DRP1 in glioma tissues,and to explore the role of DRP1 in glioma and its molecular mechanism.Methods: Expression of DRP1 was determined using tissue chip of human glioma tissues and normal brain tissues by immunohistochemistry staining.In cultured U251 glioma cells,DRP1 gene was silenced by small interfering RNA,and the proliferation of glioma cells was detected using EdU(5-Ethynyl-2’-deoxyuridine,EdU)method and CCK-8(Cell Counting Kit-8,CCK-8)method.Transwell method was used to estimate the invasion of glioma cells,while the distribution of F-actin and the morphometry of glioma cells including pseudopodia and cell fold were detected using immunofluorescence of phalloidin.The effect of DRP1 silencing on RHOA and ROCK1 expression is measured by Western blot,and the interaction between DRP1 and RHOA was identified by Co-immunoprecipitation.Results: DRP1 expression was significantly increased in glioma tissues compared to normal brain tissues,and its level was positively related to the pathological grade of glioma.Silencing DRP1 gene in cultured U251 glioma cells changed the distribution of F-actin in glioma cells,decreased the number of pseudopodia and cell fold,and inhibited the proliferation and invasion of glioma cells.Silencing DRP1 also decreased the expression of RHOA and ROCK1.The interaction between DRP1 and RHOA was also identified.Conclusions: DRP1 is expressed in human glioma tissues,and its level is positively related to the malignancy of glioma.DRP1 may promote the proliferation and invasion of glioma cells,it interacts with RHOA and may promote the proliferation and invasion of glioma cells by activating RHOA/ROCK1 pathway.