Pharmacological induction of the hypoxia response pathway in Huh7 hepatoma cells limits proliferation but increases resilience under metabolic stress

The hypoxia response pathway facilitates adaptation to low oxygen levels, primarily through hypoxia-inducible factors (HIF), which drive processes like metabolic reprogramming, erythropoiesis, angiogenesis, and tissue remodeling. This understanding has led to the successful development of HIF-inducing drugs for treating anemia, some of which are already in clinical use. However, elevated HIF levels are often linked to tumor growth, poor prognosis, and drug resistance in various cancers, including hepatocellular carcinoma (HCC). As a result, there are concerns about recommending HIF-inducing drugs in certain clinical scenarios. In this study, we examined the effects of two HIF-inducing drugs, Molidustat and Roxadustat, on the well-established HCC cell line Huh7. These drugs increased the levels of HIF-1α and HIF-2α proteins, both of which activate hypoxia response genes such as BNIP3, SERPINE1, LDHA, and EPO. Integrated transcriptomic, proteomic, and metabolomic analyses BAY 85-3934 revealed that Molidustat enhanced the expression of glycolytic enzymes, leading to a fragmented mitochondrial network and decreased cellular respiration. This metabolic shift was associated with reduced cell proliferation, a lower demand for pyrimidine supply, and an increased capacity to convert pyruvate to lactate. Additionally, cells treated with these drugs exhibited greater resistance to the inhibition of mitochondrial respiration by antimycin A, a finding that was also observed in Roxadustat-treated Huh7 cells and Molidustat-treated hepatoblastoma cells (Huh6 and HepG2). Overall, this study demonstrates that HIF-inducing drugs enhance the metabolic resilience of liver cancer cells to metabolic stress, highlighting the need for careful monitoring of patients undergoing treatment with these drugs, particularly those at risk of developing liver cancer.