ETS1 facilitates the progression of OSCC and contributes to macrophage M2 polarization via regulating NXPH4.

Oral squamous cell carcinoma (OSCC) is a common malignant tumor of the head and neck, representing a significant public health concern. Neurexophilin-4 (NXPH4) facilitates cancer cell proliferation and invasion. E26 oncogene homolog 1 (ETS1) can influence OSCC cell migration and invasion by modulating various factors. This study aims to explore the molecular mechanism of NXPH4 and ETS1 in the progression of OSCC. Bioinformatics analysis was used to analyze the NXPH4 and ETS1 expression in OSCC tissues and explore the relationship between NXPH4 and tumor stage. The mRNA and protein levels were examined by reverse transcription-quantitative polymerase chain reaction (qRT-PCR) and western blot. Besides, the OSCC tumor models were constructed, and tumor volume and weight were detected. The immunohistochemistry (IHC) assay was used to analyze Ki-67 expression in vivo. The apoptosis and reactive oxygen species (ROS) were measured by flow cytometry. The levels of malondialdehyde (MDA) and ferrous ion (Fe(2+)) were detected using the corresponding detection kits. The cell proliferation was tested by 5‑ethynyl‑2'‑deoxyuridine (EdU) staining. Tumor Immune Estimation Resource (TIMER) 2.0 and flow cytometry were used to analyze the relationship between NXPH4 and macrophage M2 polarization. The interaction between NXPH4 and ETS1 was demonstrated by chromatin immunoprecipitation (CHIP) and a dual luciferase reporter gene assay. NXPH4 expression was significantly increased in OSCC tissues and CAL-27 and SCC-25 cell lines (P < 0.01). Silencing NXPH4 inhibited the OSCC tumor growth in vivo. In human OSCC cell lines, silencing NXPH4 promoted the apoptosis, MDA, and ROS levels and suppressed the GPX4 expression (P < 0.01), cell proliferation, and macrophage M2 polarization. Furthermore, ETS1 could bind to NXPH4 and positively regulate its expression. Knockdown of ETS1 facilitated the apoptosis, increased MDA and ROS levels, and repressed the GPX4 expression, cell proliferation, and macrophage M2 polarization. However, up-regulation of NXPH4 in vitro reversed the effect of sh-ETS1 in human OSCC cell lines. ETS1 elevates the progression of OSCC and enhances macrophage M2 polarization by contributing to NXPH4. These findings identify the ETS1/NXPH4 axis as a potential therapeutic target in OSCC. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10616-025-00869-6.