In this study, we further depicted the
SLC9A1 mRNA expression pattern in different grade gliomas from CGGA and TCGA two independent databases. Higher levels of
SLC9A1 mRNA expression were observed in GBM tumors. The classical subtype is defined by the constellation of the most common genomic aberrations, such as chromosome 7 amplifications and 10 deletions, while mesenchymal subtype displayed mesenchymal differentiation [
21]. However, both classical and mesenchymal subtype are associated with worse outcome [
20,
21]. Here, we detected that
SLC9A1 mRNA expression was higher in the classical and mesenchymal molecular subtypes in CGGA dataset and
SLC9A1 mRNA expression was highest in the mesenchymal molecular subtypes in TCGA dataset. Therefore, higher
SLC9A1 mRNA expression is associated with the malignancy of gliomas. High
SLC9A1 levels of expression showed poor outcomes in each different grade gliomas. Especially, higher
SLC9A1 mRNA expression increased 1.578-fold of hazard ratio (HR), indicating as an independent factor for poor prognosis. This is consistent with the data from the Repository for Molecular Brain Neoplasia Data (REMBRANDT) with shorter overall survival in GBM patients with upregulated
SLC9A1 mRNA expression [
15]. Our immunostaining analysis of grade II–III astrocytoma and grade IV gliomas shows that NHE1 protein expression increased with increasing tumor malignancy. Taken together, our analysis (>1000 glioma cases) strongly suggests that NHE1 protein is involved in glioma tumorigenesis and progression. The underlying mechanisms could include promoting tumor proliferation and invasion [
37‐
41]. Our gene ontology analysis indicates that NHE1 protein is involved in the tumor migration and invasion by the regulation of cell adhesion and extracellular matrix organization. NHE1 protein can stimulate tumor cell proliferation via the NHE1-mediated Na
+ influx for volume increase and H
+ efflux to maintain alkaline pH
i during the cell cycle progression [
38]. Pharmacological inhibition of NHE1 has been shown to suppresses the G1/S and G2/M cell cycle phase by the reduction of cell cycle regulator expression, such as cyclin D1 and cyclin B1 [
39]. NHE1 protein also stimulates the matrix metalloproteinases family and promote tumor invasion via ERK1/2 and p38 MAPK signaling pathways [
42]. In our study, inhibition of NHE1 with HOE642 (0.5 mg/kg per day for continuous 5 days) significantly decreased the tumor volume and tumor invasion in mouse intracranial glioma animal model. HOE642 treatment caused a significant right-shift of survival curve in the glioma-bearing mice.
In both two databases, we found that
SLC9A1 mRNA expression was higher in the IDH-wildtype than in the IDH-mutant gliomas. IDH status plays an important role in the progression of gliomas and the outcome of patients [
32,
43]. In a hypoxic environment, cells rely on the reductive carboxylation of glutamine-derived α-ketoglutarate (α-KG) for lipid synthesis and reductive glutamine metabolism is also a characteristic of tumors, including gliomas [
44,
45]. The IDH enzymes play a vital role in cellular protection and in response to oxidative and energetic stress via the role of NADPH in the regeneration of the antioxidant glutathione and catalyzing the oxidative carboxylation of isocitrate to α-KG [
44,
46]. However, IDH mutant tumor cells downregulate expression of hypoxia-associated genes, such as HIF1α, and increases the risk of reactive oxygen species-mediated DNA damage [
44,
47]. NHE1 protein expression is regulated by hypoxia-inducible factor (HIF1α) and facilitates extruding H
+ to maintain Warburg effects [
15,
48]. Therefore, IDH wild-type gliomas may activate NHE1 expression to maintain pH
i in response to oxidative stress, promoting tumor cell survival.