Senegenin suppresses hepatocellular carcinoma by regulating O-GlcNAcylation

Figure 1 Expression of O-GlcNAcylation and its regulatory enzyme O-linked N-acetylglucosamine transferase in liver cancer tissues and cell lines. A: The expression of the O-GlcNAcylation regulatory enzyme O-linked N-acetylglucosamine transferase (OGT) in liver cancer tissues was detected using western blotting; B: Expression levels of O-glcnAcylated glycosylation and OGT levels in each hepatocellular carcinoma cell line by western blot. OGT: O-linked N-acetylglucosamine transferase.

Figure 2 Effects of O-GlcNAcylation and glycosylation on hepatocellular carcinoma cell proliferation and migrating. A: The impact of O-GlcNAcase inhibitors (TMG, PUG) on O-GlcNAcylation glycosylation in cells of BEL-7402 was determined using a western blot; B: The cell counting kit-8 assay was utilized to identify hepatocellular carcinoma (HCC) cell proliferating; C: Colony-forming ability of HCC cells; D: To track the migrating of liver cancer cells, the transwell assay was utilized. HCC: Hepatocellular carcinoma; OGT: O-linked N-acetylglucosamine transferase; DMSO: Dimethyl sulfoxide.

Figure 3 Effect of O-linked N-acetylglucosamine transferase overexpression on hepatocellular carcinoma cells' proliferating and migrating. A: O-linked N-acetylglucosamine transferase was overexpressed in BEL-7402 cells, and Western blot analysis was used to assess the level of cell glycosylation; B: The cell counting kit-8 assay was utilized to identify hepatocellular carcinoma (HCC) cell proliferating; C: Colony-forming ability of HCC cells; D: To find out whether liver cancer cells were migrating, transwell assay was used. OGT: O-linked N-acetylglucosamine transferase.

Figure 4 Effect of senegenin on hepatoma cells. A: Senegenin's impact on the survival rate of hepatoma cells was measured using the MTT method; B: Senegenin's impact on the growth of hepatoma cells was assessed using the cell counting kit-8 assay; C: Senegenin's ability to migrate on hepatocellular carcinoma cells was examined using the Transwell Assay. CCK8: Cell counting kit-8.

Figure 5 Effect of senegenin regulating O-GlcNAcylation glycosylation on hepatocellular carcinoma cells. A: Senegenin's impact on O-GlcNAc glycosylation and O-linked N-acetylglucosamine transferase was discovered by Western blot in hepatoma cells; B: Using the cell counting kit-8 assay, the effect of senegenin-regulated O-GlcNAc glycosylation on the proliferating of hepatocellular carcinoma cells was evaluated; C: O-GlcNAc glycosylation regulated by senegenin on colony-forming ability of hepatoma cells; D: The effect of senegenin-regulated O-GlcNAc glycosylation on the capacity of hepatocellular carcinoma cells to migrate was examined using the transwell assay.

Figure 6 Mechanism of O-GlcNAcylation glycosylation effect on hepatocellular carcinoma cells. A: Western blot analysis of endoplasmic reticulum stress protein expression levels after O-linked N-acetylglucosamine transferase (OGT) overexpression; B: Dual luciferase assay was used to measure the activity of AP-1; C: OGT overexpression results in a Western blot analysis of the levels of NF-κB pathway protein expression. ER: Endoplasmic reticulum; OGT: O-linked N-acetylglucosamine transferase; JNK: C-Jun N-terminal kinase.