Functional Role of NOXA in Hypoxia-Mediated PD-L1 Inhibitor Response in Hepatocellular Carcinoma.

Hypoxia is a crucial characteristic of hepatocellular carcinoma (HCC) and contributes to immune resistance by upregulating PD-L1 and recruiting immunosuppressive cells. However, the molecular mechanisms of hypoxia-induced immunotherapy resistance are still unclear. The hypoxia-related immunotherapy response (IRH) genes were identified and used to develop a hypoxia risk score model to predict patient survival. The model was validated using GSE233802 and EGAD00001008128 datasets. The hypoxia risk score model including NOXA effectively stratified patients based on risk and demonstrated excellent survival predictive ability ( = 0.0236). A hypoxia-induced drug-resistant (HepG2-R) cell line was established by co-culturing HepG2 cells with Jurkat T cells under CoCl-induced hypoxia and PD-L1 inhibitor administration. Prolonged exposure to hypoxia (48 h) in HepG2 cells significantly led to the increased hypoxia risk score ( < 0.02). The establishment of the HepG2-R cell line showed that prolonged hypoxia reduced cancer cell apoptosis, which implies potential treatment resistance. The effect of NOXA knockdown on the apoptosis of HepG2-R cells under the same co-culture conditions was examined. Under hypoxia and PD-L1 inhibitor treatment, NOXA knockdown increased the survival rate of HepG2-R cells and reduced early and late apoptosis. This indicates that NOXA plays a crucial role in apoptosis regulation and immune response in hypoxic tumors. NOXA knockdown significantly reduces apoptosis in immunotherapy-resistant cells induced by hypoxia. These findings provide important evidence that targeting NOXA may enhance immunotherapy efficacy and help overcome treatment resistance in HCC, highlighting its potential as a therapeutic target.