Experimental study of 3-bromopyruvate inhibiting the aerobic glycolysis of hepatocarcinoma cells

Abstract

Objective To study the effect of 3-bromopyruvate (3-BrPA) on the aerobic glycolysis pathway of hepatocarcinoma cell HepG2. Methods The human hepatocarcinoma cell HepG2 and the human normal liver cell QSG-7701 were treated with different concentration of 3-BrPA, the cell viability was detected by cell counting kit-8 (CCK-8) assay. Morphological changes of the two cells were observed under microscope. The cell apoptosis rate was detected by flow cytometry. Real-time quantitative polymerase chain reaction (FQ-PCR) and Western blotting were used for detecting the expression of c-Myc and monocarboxylate transporter 1 (MCT1) in HepG2 and QSG-7701 at mRNA and protein levels. The glucose uptake, lactate secretion and intracellular content of adenosine triphosphate (ATP) of the cells were measured by enzyme-linked method. Results The cell survival rate and the cell apoptosis rate were significantly increased with the increase of 3-BrPA concentration. Apoptosis rates of HepG2 treated with different concentrations of 3-BrPA (25, 50, 75 μmol/L) were (4.64±2.68)%, (14.57±8.41)%, (40.55±23.41)% respectively, the apoptosis rate of control group (0 μmol/L) was (5.62±3.24)%, and the concentration of 75 μmol/L was significantly different from that of the control group (P=0.025). Compared with QSG-7701, the expression of c-Myc and MCT1 in HepG2 were significantly higher than those in QSG-7701 at mRNA and protein levels (P=0.001, 0.000), and decreased when the concentration of 3-BrPA was 75 μmol/L (P=0.041, 0.000). The results of glucose metabolism analysis showed that the glucose uptake, lactate secretion and intracellular ATP of HepG2 decreased gradually when the concentration of 3-BrPA was 75 μmol/L (P=0.035, 0.000, 0.045), but no significant change in QSG-770 (P=0.898, 0.841, 0.112). Conclusion 3-BrPA can inhibit the growth of HepG2 and promote the cell apoptosis by inhibiting the expression of c-Myc/MCT1 and the aerobic glycolysis of HepG2. Key words: 3-bromopyruvate; Hepatocellular carcinoma; c-Myc; Monocarboxylate transporter 1; Aerobic galycolysis