BMP4 reprograms glucose metabolism in hepatocellular carcinoma (HCC) cells by upregulating glycogen synthesis through the SMAD/SLC2A1 (GLUT1) signaling axis

Abstract

Abstract Background Excessive hepatic glycogen accumulation benefits tumorigenesis and cancer cell survival. We previously reported that BMP4 was elevated in Hepatocellular carcinoma (HCC) and augmented HCC cell survival under hypoxia and hypoglycemia conditions by promoting the glycolysis pathway. However, the mechanism underlying BMP4’s effect on glucose metabolism remains elusive. In this study, we investigated the effect of BMP4 on hepatic glycose metabolism through glucose transporter SLC2A1 (GLUT1) in HCC cells. Methods The expression of BMP4 and SLC2A1 were acquired by analyzing the TCGA-LIHC dataset, as well as by immunohistochemical analysis of the 40 pairs of human HCC samples and para-tumor tissues. Gene expressions were detected by qPCR, immunoflurorescence staining, and Western blotting. Overexpression and silencing of BMP4 were accomplished through adenoviruses Ad-B4 and Ad-siB4 infection. Hepatic glycogen was detected by PAS staining. SLC2A1(GLUT1) function was blocked by the inhibitor BAY-876. ChIP assay was used to determine the binding of SMADs to the promoter region of SLC2A1 in HCC cells. Lastly, the in vivo effect of BMP4-regulated SLC2A1 on HCC tumor growth was assessed in a xenograft model of HCC. Results The elevated expression of BMP4 in HCC tumor tissues was highly correlated with hepatic glycogen accumulation in clinical samples. SLC2A1 was highly expressed in HCC tumor tissue and correlated with clinical stage and prognosis. Exogenous BMP4 augmented glycogen accumulation and up regulated the expression of glycogen synthesis-related genes in Huh7 and HepG2 cells, both of which were effectively blunted by SLC2A1inhibitor BAY-876. In mechanism, BMP4 activated SMAD5 to regulate the promoter of SLC2A1to enhance its expression. The in vivo xenograft experiments revealed that BMP4 promoted glycogen accumulation and tumor growth, which were effectively diminished by BAY-876. Conclusion These results demonstrate that BMP4 can reprogram hepatic glycogen metabolism and promote tumor growth of HCC cells through SMAD/SLC2A1 axis, which may be exploited as novel therapeutic targets for HCC treatment.