Mechanism of HAND2 Regulating Glycolytic Metabolism and Metastasis Through TLR4/NOX2/DUOX2 Signaling Axis in Cervical Cancer

ABSTRACT Objective Cervical cancer (CC) ranks among the common malignant tumors. HAND2 is actively expressed in healthy endometrium, and its expression level correlates with the malignancy of uterine diseases. Methods The GEO dataset GSE7410 was obtained and analyzed for gene expression, and differentially expressed genes were screened using GEO2R. The expression of HAND2 in CC and normal tissues was determined using the GEPIA database. SiHa cell viability, proliferation, migratory, and invasive potential were evaluated. Glucose uptake, lactate production, and ATP levels were measured in SiHa cells. To analyze apoptosis and the cell cycle in SiHa cells, flow cytometry was performed. Western blot was performed to detect glycolysis‐related genes LDHA, HK2, and PKM2, and EMT‐related genes E‐cadherin and N‐cadherin in SiHa cells. A dual luciferase reporter assay was performed to detect the regulation of TLR4, NOX2, and DUOX2 transcription by HAND2. SiHa cells stably overexpressing HAND2 were injected into nude mice to construct CC nude mouse model and metastasis models. Tumor histopathology and metastasis were assessed. Results HAND2 was down‐regulated in CC, and its low expression level correlated with poor prognosis in CC patients. HAND2 indirectly inhibited NOX2/DUOX2 activity through the TLR4 pathway, suppressing glycolytic metabolism, cell proliferation, and migration ability. In the tumor microenvironment, aberrant expression of NOX2 and DUOX2 led to elevated ROS levels, which in turn affected CC cell apoptosis and cell cycle. Conclusion The TLR4/NOX2/DUOX2 axis mediates HAND2 regulation of CC cell glycolytic metabolism, as well as cell proliferation and migration capacity.