Sialic Acid-Modified O‑GlcNAc Transferase Inhibitor Liposome Presents Antitumor Effect in Hepatocellular Carcinoma

O-linked-N-acetylglucosaminylation (O-GlcNAcylation) plays a key role in hepatocellular carcinoma (HCC) development, and the inhibition of O-GlcNAcylation has therapeutic potential. To decrease the systemic adverse events and increase targeting, we used sialic acid (SA)-decorated liposomes loaded with OSMI-1, an inhibitor of the O-GlcNAcylation, to further improve the anti-HCC effect. Fifty pairs of HCC tissue samples and the cancer genome atlas database were used to analyze the expression of O-GlcNAc transferase (OGT) and its effects on prognosis and immune cell infiltration. OSMI-1 cells were treated with SA and liposomes. Western blotting, immunofluorescence, cell proliferation assay, flow cytometry, enzyme-linked immunosorbent assay, immunohistochemistry, and tumorigenicity assays were used to investigate the antitumor effect of SA-modified OSMI-1 liposomes in vitro and in vivo. OGT was highly expressed in HCC tissues, negatively correlated with the degree of tumor infiltration of CD8+ and CD4+T cells and prognosis, and positively correlated with the degree of Treg cell infiltration. SA-modified OSMI-1 liposome (OSMI-1-SAL) was synthesized with stable hydrodynamic size distribution. Both in vitro and in vivo, OSMI-1-SAL exhibited satisfactory biosafety and rapid uptake by HCC cells. Compared to free OSMI-1, OSMI-1-SAL had a stronger capacity for suppressing the proliferation and promoting the apoptosis of HCC cells. Moreover, OSMI-1-SAL effectively inhibited tumor initiation and development in mice. OSMI-1-SAL also promoted the release of damage-associated molecular patterns, including anticalreticulin, high-mobility-group protein B1, and adenosine triphosphate, from HCC cells and further promoted the activation and proliferation of the CD8+ and CD4+T cells. In conclusion, the OSMI-1-SAL synthesized in this study can target HCC cells, inhibit tumor proliferation, induce tumor immunogenic cell death, enhance tumor immunogenicity, and promote antitumor immune responses, which has the potential for clinical application in the future

MOESM1 of YAP promotes multi-drug resistance and inhibits autophagy-related cell death in hepatocellular carcinoma via the RAC1-ROS-mTOR pathway

Additional file 1: Figure S1. (A) The efficiency of YAP knockdown in BEL/FU cells. (B) The protein levels of cleaved PARP and cleaved caspase-3 were detected in BEL/FU cells with or without YAP knockdown after treatment with 5-Fu or DOX for 48 h by western blot. The protein amount of cleaved PARP and cleaved caspase-3 were measured and quantified by analysis of densitometry. (C) The efficiency of ATG5 and BECN1 knockdown in BEL/FU cells. (D) The mRNA expression of YAP in HCC cell lines and the protein expression of YAP in HCC-LM3 and SK-Hep1 cells. (E) The protein level of the autophagy marker LC3B was measured in BEL/FU cells with or without treatment with rapamycin (20 nM) for 6 h by western blot analysis. (F) The expression of p-mTOR, p-S6 and 8-OHdG was examined by IHC analysis of xenograft tumour tissues from Balb/c nude mice treated with verteporfin and DOX (scale bar: 50 µm/25 µm). (G) After treatment with 5-Fu and DOX, the amount of apoptosis markers, cleaved PARP and cleaved caspase-3, in BEL/FU cells with or without treatment with rapamycin for 48 h was measured by western blot. The protein amount of cleaved PARP and cleaved caspase-3 were measured and quantified by analysis of densitometry. Data are presented as the mean ± SD. *p < 0.05, **p < 0.01

MOESM2 of YAP promotes multi-drug resistance and inhibits autophagy-related cell death in hepatocellular carcinoma via the RAC1-ROS-mTOR pathway

Additional file 2: Figure S2. (A-D) The protein amount of LC3B-II/LC3B-I was measured and quantified in HCC cell lines (BEL/FU, SK-Hep1, BEL-7402, and HCC-LM3) with YAP overexpression or knockdown under Earle’s Balanced Salt Solution (EBSS) starvation conditions in the presence or absence of chloroquine (CQ). (E–G) The protein amount of p-mTOR, p-S6 and p-4E-BP1 was measured and quantified in HCC cell lines (BEL/FU, SK-Hep1, BEL-7402, and HCC-LM3) with YAP overexpression or knockdown. (I, J) The protein amount of p-mTOR, p-S6 and p-4E-BP1 was measured and quantified in BEL/FU cells with verteporfin treatment and BEL/FU cells with or without YAP knockdown after treatment with NAC. CM: complete medium. Data are presented as the mean ± SD. *p < 0.05, **p < 0.01, ns, no significance