RESUMEN
BACKGROUND: Gliomas are the most common primary malignant tumours of the central nervous system, and neddylation may be a potential target for the treatment of gliomas. Our study analysed neddylation's potential role in gliomas of different pathological types and its correlation with immunotherapy. METHODS: Genes required for model construction were sourced from existing literature, and their expression data were extracted from the TCGA and CGGA databases. LASSO regression was employed to identify genes associated with the prognosis of glioma patients in TCGA and to establish a clinical prognostic model. Biological changes in glioma cell lines following intervention with hub genes were evaluated using the CCK-8 assay and transwell assay. The genes implicated in the model construction were validated across various cell lines using Western blot. We conducted analyses to examine correlations between model scores and clinical data, tumor microenvironments, and immune checkpoints. Furthermore, we investigated potential differences in molecular functions and mechanisms among different groups. RESULTS: We identified 249 genes from the Reactome database and analysed their expression profiles in the TCGA and CGGA databases. After using LASSO-Cox, four genes (BRCA1, BIRC5, FBXL16 and KLHL25, p < 0.05) with significant correlations were identified. We selected FBXL16 for validation in in vitro experiments. Following FBXL16 overexpression, the proliferation, migration, and invasion abilities of glioma cell lines all showed a decrease. Then, we constructed the NEDD Index for gliomas. The nomogram indicated that this model could serve as an independent prognostic marker. Analysis of the tumour microenvironment and immune checkpoints revealed that the NEDD index was also correlated with immune cell infiltration and the expression levels of various immune checkpoints. CONCLUSION: The NEDD index can serve as a practical tool for predicting the prognosis of glioma patients, and it is correlated with immune cell infiltration and the expression levels of immune checkpoints.
Asunto(s)
Biomarcadores de Tumor , Neoplasias Encefálicas , Regulación Neoplásica de la Expresión Génica , Glioma , Humanos , Glioma/genética , Glioma/inmunología , Glioma/patología , Pronóstico , Línea Celular Tumoral , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/inmunología , Neoplasias Encefálicas/patología , Biomarcadores de Tumor/genética , Biomarcadores de Tumor/metabolismo , Proteína NEDD8/genética , Proteína NEDD8/metabolismo , Microambiente Tumoral/genética , Microambiente Tumoral/inmunología , Proteínas de Punto de Control Inmunitario/genética , Proteínas de Punto de Control Inmunitario/metabolismo , Proliferación Celular/genética , Proteínas F-Box/genética , Proteínas F-Box/metabolismo , Femenino , Perfilación de la Expresión Génica/métodos , Bases de Datos Genéticas , Movimiento Celular/genética , MasculinoRESUMEN
BACKGROUND: Intracerebral hemorrhage (ICH), a subtype of devastating stroke, carries high morbidity and mortality worldwide. CircRNA AFF2 (circAFF2) was significantly increased in ICH patients, but the underlying mechanism of circAFF2 is unknown. METHODS: Hemin was employed to treat neuronal cells to mimic ICH in vitro. Mice were injected with collagenase VII-S to establish in vivo ICH models. Genes and protein expressions were detected using qRT-PCR and Western blotting. The interaction among circAFF2, miR-488, and CLSTN3 was validated by dual-luciferase reporter assay and RNA-RIP. Cell viability, MDA, iron, GSH, and lipid ROS were examined using the MTT, the commercial kits, and flow cytometry, respectively. ICH injury in mice was evaluated using neurological deficit scores and brain water measurements. RESULTS: CircAFF2 was significantly increased in ICH in vivo and in vitro models. CircAFF2 bound to miR-488 and knockdown of circAFF2 or overexpression of miR-488 inhibited hemin-induced injury of neuronal cells as indicated by increased cell viability and reduced markers of oxidative stress and lipid peroxidation. CLSTN3 was the downstream target of miR-488. Silencing of circAFF2 or miR-488 overexpression reduced CLSTN3 expression and protected against the injury of neuronal cells. In vivo experiments finally confirmed that circAFF2 knockdown attenuated mice ICH injury via the miR-488/CLSTN3 axis. CONCLUSION: CircAFF2 promotes the injury of neuronal cells and exacerbates ICH via increasing CLSTN3 by sponging miR-488, suggesting that circAFF2 may be a potential therapeutic target for ICH treatment.
Asunto(s)
Lesiones Encefálicas , MicroARNs , Animales , Humanos , Ratones , Encéfalo/metabolismo , Lesiones Encefálicas/metabolismo , Proteínas de Unión al Calcio/metabolismo , Hemorragia Cerebral/metabolismo , Hemina/farmacología , Hemina/metabolismo , Proteínas de la Membrana/metabolismo , MicroARNs/genética , MicroARNs/metabolismoRESUMEN
Fyn-related kinase (FRK), a member of the Src-related tyrosine kinase family, functions as a tumor suppressor in several malignancies. We previously showed that FRK overexpression inhibited the growth of glioma cells. However, it is unknown whether FRK is equally effective against intracranial glioma in vivo, and the mechanism by which FRK influences glioma cell growth remains unclear. In this study, we found that tumor volume was reduced by about one-third in mice with FRK overexpression, which showed improved survival relative to controls. Immunofluorescence analysis revealed that FRK overexpression inhibited glioma cell proliferation and induced their apoptosis. Importantly, in vitro we further found that FRK decreased the expression of integrin subunit ß1 (ITGB1) at both the mRNA and protein levels. FRK also inhibited transactivation by ITGB1, resulting in the suppression of its target proteins AKT and focal adhesion kinase (FAK). ITGB1 overexpression promoted glioma cell growth and partially reduced FRK-induced growth suppression. These results indicate that FRK inhibits human glioma growth via regulating ITGB1/FAK signaling and provide a potential therapeutic target for the treatment of glioma.