Extracellular Citrate Treatment Induces HIF1α Degradation and Inhibits the Growth of Low-Glycolytic Hepatocellular Carcinoma under Hypoxia

Abstract

Simple Summary Patients with low-glycolytic hepatocellular carcinoma (HCC) show better clinical outcomes than those with hypoxic and high-glycolytic HCC. Low-glycolytic HCCs seem to utilize carbon sources other than glucose for metabolic fuel and tumor growth. However, by increasing tumor size, its outgrowth perfusion generates hypoxic foci inside the tumor and becomes more aggressive and resistant to therapy. In this study, we found that SLC13A5/NaCT is an important solute carrier (SLC) in low-glycolytic HCCs. To adapt to hypoxic conditions, low-glycolytic cancer cells have to switch metabolism from oxidative phosphorylation to hypoxia-induced glycolysis by the upregulation of HIF1 alpha. However, extracellular citrate treatment in HCCs with high SLC13A5/NaCT expression had reduced glucose uptake due to HIF1 alpha degradation, inducing the failure of metabolic adaptation to hypoxia, resulting in anti-cancer effects in in vitro and in vivo animal models. HCC is well known for low glycolysis in the tumors, whereas hypoxia induces glycolytic phenotype and tumor progression. This study was conducted to evaluate the expression of SLCs in human HCCs and investigated whether extracellular nutrient administration related to SLCs in low-glycolytic HCC can prevent hypoxic tumor progression. SLCs expression was screened according to the level of glycolysis in HCCs. Then, whether extracellular nutrient treatment can affect hypoxic tumor progression, as well as the mechanisms, were evaluated in an in vitro cell line and an in vivo animal model. Low-glycolytic HCCs showed high SLC13A5/NaCT and SLC16A1/MCT1 but low SLC2A1/GLUT1 and HIF1 alpha/HIF1 alpha expression. Especially, high SLC13A5 expression was significantly associated with good overall survival in the Cancer Genome Atlas (TCGA) database. In HepG2 cells with the highest NaCT expression, extracellular citrate treatment upon hypoxia induced HIF1 alpha degradation, which led to reduced glycolysis and cellular proliferation. Finally, in HepG2-animal models, the citrate-treated group showed smaller tumor with less hypoxic areas than the vehicle-treated group. In patients with HCC, SLC13A5/NaCT is an important SLC, which is associated with low glycolysis and good prognosis. Extracellular citrate treatment induced the failure of metabolic adaptation to hypoxia and tumor growth inhibition, which can be a potential therapeutic strategy in HCCs.