The Function And Mechanism Of ANP32A In Glioma Development

Background and ObjectiveGlioma is the most common primary malignant tumor of the central nervous system.As diffusely infiltrating growth and invading normal brain tissue,gliomas are often difficult to excise completely by surgery,and cure by postopeirative radiotherapy and chemotherapy.Surgery,radiotherapy,and temozolomide(TMZ)chemotherapy are first-line treatment for newly diagnosed glioma patients,but the therapeutic effects are not satisfactory.Acid-rich nuclear phosphoprotein 32(ANP32)family membersare closely associated with tumor progression,injury protection,and drug tolerance.Phosphoprotein 32 A(ANP32A),the earliest discovered and most studied ANP32 family protein,is closely related to the progression of various malignant tumors such as liver cancer,prostate cance,breast cancer and colorectal cancer.Our previous studies have shown that ANP32A is highly expressed in mammalian brain tissue and participates in neurodevelopmental processes.Further studies have shown that ANP32A is highly expressed in glioma tissues.However,the role and mechanism of ANP32A in the development of glioma is still unclearMethodsLentiviral vector carrying human ANP32A sequence or its interference sequence was constructed,lentivirus was prepared and infected human glioma cell line U251,and ANP32A high or low expression U251 cell lines were screened with puromycin;compared with control cells,the effect of high or low expression of ANP32A on U251 was observed,including cell proliferation and apoptosis determined using CCK-8 assay and flow cytometry;wound healing and Transwell assay determine U251 cell migration and invasion;the expression of cellular protein was determined using immunocytochemical staining and Western blot.The effect of ANP32A on the tumorigenesis and migration of U251 cells was determined using zebrafish xenograft model.Immunohistochemistry was employed to analyze the expression of ANP32A in glioma tissues from patients,and Kaplan Meier method was used to analyze the correlation between ANP32A expression and patient prognosis.Results1.The ANP32A high or low expression human glioma U251 cell lines were established;ANP32A high or low expression can promote or inhibit U251 cell proliferation,migration and invasion,and inhibit or promote U251 cell apoptosis2.ANP32A could induce the expression of HuR,SET,Vimentin and Tublin in U251 cells,and suppress the PP2A expression.Meanwhile,ANP32A could inhibit the injury induced by hypoxia and chemical drugs in U251 cells.3.High or low ANP32A expression could activate or inhibit Akt phosphorylation in U251 cells,and extracellular stimulation of human recombinant protein ANP32A can activate the Akt signaling pathway in U251 cells in a time-dependent manner.4.In zebrafish xenografts,high or low ANP32A expression could promote or inhibit tumor growth and migration.5.The expression level of ANP32A was positively correlated with the pathological stage of glioma,and the expression level of ANP32A was higher in the pathological tissues of high-grade glioma compared with low-grade glioma.High ANP32A expression is an independent predictor of poor prognosis in patients.The overall survival rate of patients with high ANP32A expression glioma is significantly lower than patients with low ANP32A expressionConclusionThe results showed that ANP32A promote the progression of glioma.The mechanism may be related to the regulation of the expression of protein factors such as ANP32A regulating RNA binding protein HuR and activation of Akt signaling pathway.High ANP32A expression is an independent predictor of poor prognosis in patients.This study suggests that ANP32A could be served as a marker for molecular diagnosis and prognosis evaluation in glioma,providing new ideas and therapeutic targets for the diagnosis and treatment of glioma.