A novel L-shaped ortho-quinone analog and 2-Deoxy-D-Glucose a synergistic approach to inhibit hepatocellular carcinoma cell proliferation.

2-Dexy-D-glucose (2-DG) is a chemotherapy drug that blocks the glycolytic process, thereby exerting its metabolism inhibitor and tumor growth inhibitor function in hepatocellular carcinoma (HCC) cells. However, 2-DG requires prolonged high-dose treatment to achieve enough therapeutic efficacies, which may lead to persistent side effects such as elevated blood glucose levels, dizziness, and nausea. We have screened a novel L-shaped ortho-quinone analog named TC1, which exhibits excellent anti-tumor activity with broad-spectrum cytotoxicity against multiple tumor cell lines. In the meanwhile, it also shows a strong potential to inhibit metabolism in tumor cells, which makes it a good candidate for combined treatment with 2-DG. We used combined treatment of TC1 and 2-DG on HCC cell lines and analyzed cell proliferation, apoptosis, migration, invasion, and mitochondrial function. Additionally, we examined the effects of this combination therapy on nude mice bearing transplanted tumor. Furthermore, we explored the potential mechanisms using RNA sequencing and verified by RT-qPCR and immunofluorescence staining. In vitro experiments showed that the combination treatment of TC1 and 2-DG synergistically inhibited the proliferation of HCC cell lines including SMMC-7721 cells and Hepa1-6 cells, induced apoptosis, depleted ATP content in cells and increased ROS content in cells. In vivo experiments showed the combination treatment effectively inhibited the growth of SMMC-7721 cell transplanted tumors and prolonged the survival time of mice. Through RNA sequencing, we found that MAPK4 was down-regulated by the combination treatment. Through real-time quantitative PCR and immunofluorescence staining, we confirmed that both MAPK4 expression and AKT phosphorylation were significantly decreased, together with the increasing expression of BAX in the combination treatment group. The combination treatment significantly inhibits HCC cell proliferation both in vitro and in vivo, suggesting a potential new strategy for HCC treatment.