Mechanism Of PDGFRβ Regulating Aerobic Glycolysis In Osteosarcoma Cells

Objectives: To study the correlation between platelet-derived growth factor receptor-beta(PDGFRβ)and the aerobic glycolysis of osteosarcoma cell line HOS and its regulatory mechanism on aerobic glycolysis.Methods: The expression profiles were downloaded from the GEO databases,and then GSEA,Pearson’s correlation test,and PPI correlation analysis were used to evaluate the enrichment of glucose metabolism in osteosarcoma and the relationship between PDGFRβ and glycolysis-targeted genes.Then we confirmed the regulation and mechanism of glycolysis-related molecules by PDGFRβ using q RT-PCR and western blotting in vitro.The glucose consumption and lactate production assays were performed to detect glucose metabolism in osteosarcoma cells.Additionally,changes in mitochondrial membrane potential were observed using a fluorescence microscope.Subsequently,after treating HOS cells with PDGFBB or PI3 K pathway inhibitor LY294002 or MEK pathway inhibitor U0126 or Warburg effect inhibitor DCA,western blotting and glucose metabolism experiments were used to detect the specific mechanism of PDGFRβ regulating aerobic glycolysis of osteosarcoma.Results: 1.Bioinformatics analysis showed that compared with adjacent tissues,glycolysis gene set was significantly enriched in osteosarcoma,and PDGFRβ was significantly positively correlated with the expression of some glycolytic target genes in osteosarcoma.2.After inhibiting the expression of PDGFRβ,the results of q RT-PCR and western blotting showed that the expression levels of key glycolytic molecules such as GLUT1,PDK1,PFK1,PKM2 and LDHA in HOS cells decreased.The protein expression level of HK2 decreased,but there was no significant difference in m RNA expression level.3.After inhibiting PDGFRβexpression,the glucose consumption experiment and lactate production experiment of HOS cells showed that the consumption of glucose by the cells was significantly reduced,the accumulation of extracellular glucose increased,and the production of lactate was significantly reduced.4.After inhibiting the expression of PDGFRβ,MMP decreased significantly.5.After activating PDGFRβ with PDGFBB,the expression levels of key glycolytic molecules GLUT1,PDK1,PFK1,PKM2,LDHA,and HK2 in HOS cells gradually increase over time.6.After the cells were treated with PDGFBB,the glucose consumption and lactate production experiments of HOS cells showed that the glucose consumption of the cells increased significantly and the extracellular glucose accumulation decreased.The production of lactic acid increased significantly.7.After treating the cells with PDGFBB,the MMP in HOS cells gradually increased with the increase of stimulation time.8.After inhibiting the expression of PDGFRβ,western blotting showed that the expression of c-Myc in the cell and the nucleus was decreased,and after the cells were treated with PDGFBB,the expression of the transcription factor c-Myc was increased in a time-dependent manner.9.The cells were treated with PDGFBB,PDGFRβinhibitor Sunitinib,PI3 K pathway inhibitor LY294002,MEK pathway inhibitor U0126,and Warburg effect inhibitor DCA for 24 hours.Western blotting results showed that PDGFBB /PDGFRβ mediated the expression of c-Myc by activating PI3 K /Akt /m TOR pathway,and then promoted the aerobic glycolysis of osteosarcoma cells.Conclusions: This work validated that PDGFRβ has a significant positive regulatory effect on aerobic glycolysis of osteosarcoma cells,and the aerobic glycolysis triggered by PDGFRβ is not secondary to the impaired mitochondrial functional state in the cell.The PDGF/PDGFRβsignal mainly regulates the aerobic glycolysis of osteosarcoma cells through the PI3K/AKT/m TOR/c-Myc pathway.