IRF2/SRD5A3 Promotes Malignant Growth and Glycolysis of Glioma Cells by Regulating ERK/CREB Pathway

Abstract

Background: Glioma is a common and aggressive primary malignant brain tumor. However, the occurrence and development of glioma has not been well reported. Our study aims to detect the function and mechanism of steroid 5 alpha-reductase 3 (SRD5A3) in the proliferation and metabolism of glioma cells. Methods: SRD5A3 expression was estimated by bioinformatics analysis and western blot assay. Next, U118MG and LN229 cells were separated into si-control, si-SRD5A3-1 and si-SRD5A3-2 groups, respectively. Cell growth ability was determined by cell counting kit-8 and colony formation assays, whereas its mobility and cycle were measured by Transwell and western blot assays. The metabolism of glioma was determined by the corresponding enzyme-linked immuno sorbent assay kit and Seahorse machine. Subsequently, the transcriptional factor of SRD5A3 was predicted by University of California Santa Cruz (UCSC) and PROMO database and confirmed by dual luciferase assay and chromatin immunoprecipitation-polymerase chain reaction (ChIP-PCR) assay. The protein pattern of extracellular regulated protein kinase (ERK)/cAMP response element binding protein (CREB) pathway related molecules was examined by western blot. Results: SRD5A3 expression was significantly increased in glioma samples, p < 0.05. Functionally, U118MG and LN229 cells in si-SRD5A3-1 or si-SRD5A3-2 group revealed the obvious reduction in its growth, cycle, mobility and metabolism in contrast with those in the si-control group, p < 0.01. Moreover, transcriptional factor interferon regulatory factor 2 (IRF2) bound to SRD5A3 and affected its expression in glioma cells. Furthermore, si-SRD5A3 antagonized the function of IRF2 overexpression in modulating glioma cell proliferation, mobility, cycle and metabolism. The p-ERK and p-CREB levels were obviously increased after IRF2 upregulation, whereas the addition of si-SRD5A3 antagonized the effect of IRF2 overexpression on p-ERK and p-CREB levels, p < 0.01. Conclusions: In summary, the findings of in vitro studies insinuated that the administration of si-SRD5A3 suppressed the growth, mobility and metabolism of glioma cells, and SRD5A3 was transcriptionally regulated by IRF2.