| Object Glioma is a primary malignant tumor of the central nervous system(CNS),accounting for 80% of the malignant tumors in the brain.Diffuse low grade gliomas(DLGG)is a special type of glioma,pathologically including Ⅱ and Ⅲ astrocytoma,and fewer oligodendrocytes tumor astrocytoma.Different from the general low-grade glioma,the growth of DLGG is progressive and invasive,and it will inevitably turn into the high-grade glioma eventually.Therefore,the mechanism of the invasive growth of DLGG is crucial for the treatment.Previous studies have shown that myelin binding to NgR can regulate glioma growth,but the regulation of DLGG is not clear.This study aims to study the mechanism of myelin binding to NgR to regulate DLGG invasion and migration through the PI3K/AKT/GSK3 /Rac-1 pathway,so as to provide new ideas for the diagnosis and treatment of DLGG.Materials and methods1.The expression of NgR,GSK3β,Rho A and Rac-1 in 69 NDLGG samples and 50 DLGG samples were determined by RT-PCR.2.DLGG specimens were divided into two parts.One part was immunohistochemical analysis of the expression of NgR,GSK3β,Rho A and Rac-1,and the other part was extraction of DLGG primary cells and transfection of virus to knock down and overexpress NgR.3.To cultivate primary cell expressed in PCR tested and knock NgR expression after low situation,and carries on the transwell migration and invasion experiment and control group,and use access kit predict pathways involved,and western blot test-pathway expression,use after testing the expression of downstream signal pathway inhibitors,CO-IP test combined with channel signal control targets4.The primary cells were injected into the intracranial of mice,and the growth of intracranial tumors was detected by imaging in vivo,and the photos were taken and recorded.Results1.PCR showed that NgR,GSK3β,Rho A,Rac-1 were expressed in NDLGG and DLGG.In addition,the expression of NgR,GSK3β,Rac-1 in DLGG was significantly higher than that in NDLGG,while the expression of Rho A in NDLGG was significantly higher than that in DLGG2.When DLGG was cultured in a complete medium coated with nogo-66 or MAG,its migration and invasion ability was significantly stronger than that of DLGG cells without nogo-66 or MAG,and the migration and invasion ability of DLGG was significantly reduced and enhanced when NgR was down-regulated and overexpressed.3.The changes of the downstream signaling pathway were detected after NgR knockdown.The PI3K/AKT signaling pathway was significantly blocked after NgR knockdown in primary DLGG cells,and the major molecules in the PI3K/AKT signaling pathway were blocked after NgR knockdown by WB.After pathway inhibition,the expression of downstream signal GSK3β was obviously blocked.The co-ip shows that Rac-1 is the downstream regulator of GSK3βThe results of animal model experiments showed that the overexpression of NgR could significantly promote the intracranial growth of DLGG,and the overexpression of NgR could significantly promote the proliferation and invasion of DLGG in situ.Conclusions The above experimental results indicated that NgR regulates Rac-1 through the PI3K/AKT/GSK3β signaling pathway to promote the migration and invasion of DLGG.This study preliminarily revealed the molecular mechanism of ngr-mediated DLGG invasion and migration,providing a new direction for the treatment of DLGG in the future... |
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