The Mechanism Underlying The Role Of ACC1 Knockdown In The Migration And Invasion Of Glioma Cells

BackgroundGlioma is a common malignant tumor of the central nervous system which has a high incidence,a high degree of malignancy,complicated treatment and a poor prognosis.The tumors infiltrate diffusely into regions of the normal brain rendering total surgical extirpation impossible and effective local radiotherapy difficult.Changes in tumor cell metabolism and related metabolic enzymes have been shown to be closely related to tumorigenesis and development.Although the TCGA database shows that the expression level of Acetyl-CoA carboxylase 1(ACC1)mRNA in glioma is significantly lower than that in normal brain tissue,it has not been reported how ACC1 affects the occurrence and development of glioma.This study aimed to explore the effect of ACC1 on the migration and invasion of glioma cells and its possible mechanism,and to provide an comprehensive theoretical basis for the relationship between energy metabolism of glioma cells and tumorigenesis and development.ObjectiveIn this study,by knocking down ACC1 in glioma cell U251,we clarified the effect of ACC1 on glioma cell migration and invasion,providing a new perspective for understanding the relationship between the energy metabolism of cells and the occurrence and development of glioma.MethodsU251 cell were used in this study.Knockdown of ACC1 in U251 cell was performed by lentiviral transfection technology.The effects of knockdown of ACC1 on cell migrationand invasion were investigated by Transwell migration/invasion assay and scratch test.Western blot was used to detect the expression of migration and invasion proteins Vimentin,Fibronectin and uPA.Then,ROS level in the cells was detected by flow cytometry.N-Acetyl-L-cysteine(NAC),an antioxidant with free radical scavenging function,was used to verify the relationship between ROS and changes in migration and invasion of U251 cell after knocking down ACC1.Finally,through the treatment of related inhibitors,the signaling pathways that affect the glioma cells migration and invasion ability due to the increase of ROS level caused by knockdown of ACC1 were explored.ResultsKnockdown of ACC1 increased migration and invasion of U251 cell.Western blot results showed that the expression level of ACC1,Vimentin,Fibronectin,uPA and H3K9 ac was increased in ACC1-knockdown U251 cell.Flow cytometry was used to detect the level of ROS.The results showed that knockdown of ACC1 increased the level of ROS in U251 cell.Then,the correlation between ROS and glioma cells migration and invasion was verified.We found that the antioxidant NAC reduced the level of ROS in U251 cell and inhibited knockdown of ACC1-induced mediated migration and invasion of U251 cell.Western blot results showed that the expression of related proteins Vimentin,Fibronectin and uPA were decreased after NAC treatment.Moreover we found that the level of p-ERK1/2 was increased in ACC1-knockdown U251 cell.Through the application of MEK/ERK inhibitor U0126,it was found that U0126 inhibited the U251 cell migration and invasion drivend by knocking down ACC1.This indicates that elevated ROS may promote the migration and invasion of U251 cell through ERK1/2 signaling pathway.ConclusionsKnockdown of ACC1 promotes glioma cell migration and invasion by ROS-ERK1/2signaling pathway.