Table 1_NFYC upregulates KLF1 expression and activate LDHA to drive glycolysis and tumor growth in glioblastoma cells.docx

ObjectiveMetabolic reprogramming is a hallmark of glioblastoma multiforme (GBM), with lactate dehydrogenase A (LDHA) playing a key role in aerobic glycolysis. However, the upstream transcriptional mechanisms driving LDHA overexpression in GBM remain poorly understood. This study aims to investigate the transcriptional regulatory network governing LDHA-mediated glycolysis and to explore the functional roles of KLF1 and NFYC in GBM progression.
MethodsBioinformatic analysis of The Cancer Genome Atlas data was performed to assess LDHA, KLF1, and NFYC expression in glioma. glioblastoma multiforme cell lines were used for loss- and gain-of-function studies by siRNA/shRNA knockdown and overexpression, including assessments of glycolytic flux, mitochondrial metabolism, cell proliferation, and apoptosis Metabolic activity was assessed using Seahorse extracellular flux analysis. Transcriptional regulation was evaluated by dual-luciferase reporter and chromatin immunoprecipitation (ChIP) assays. Tumor growth was assessed in a subcutaneous xenograft model.
ResultsLDHA was significantly upregulated in GBM and associated with poor prognosis. LDHA knockdown suppressed tumor growth, glycolysis, mitochondrial respiration, and induced apoptosis. KLF1 was identified as a direct transcriptional activator of LDHA. NFYC was shown to bind the KLF1 promoter and positively regulate its expression. Functional studies demonstrated that the NFYC-KLF1-LDHA axis promotes GBM cell proliferation, inhibits apoptosis, and enhances glycolytic and mitochondrial metabolism. The oncogenic effects of NFYC were partially reversed by KLF1 knockdown, and vice versa.
ConclusionThis study reveals a novel hierarchical transcriptional pathway in which NFYC regulates KLF1, which in turn activates LDHA, driving aerobic glycolysis and tumor progression in GBM. Targeting the NFYC-KLF1-LDHA axis may represent a promising therapeutic strategy for glioblastoma.