BACKGROUND:Liver cancer is the fifth leading cause of cancer death worldwide, but early diagnosis and treatment of liver cancer remains a clinical challenge. How to screen and diagnose liver cancer early and prolong the survival rate is still the focus of researchers.METHODS:Cell experiments were used to detect the effect of WZ35 on the colony formation ability and proliferation activity of hepatoma cells, nude mouse experiment to observe the in vivo anticancer activity and toxic side effects of WZ35; metabolomics analysis, glucose metabolism experiment and Seahorse analysis of liver cancer cells treated with WZ35; cell experiments combined with bioinformatics analysis to explore the mechanism of WZ35-mediated metabolic reprogramming to exert anticancer activity; tissue microarray and case analysis to evaluate the clinical significance of biomarkers for early diagnosis, treatment and prognosis evaluation of liver cancer.RESULTS:WZ35 inhibited the proliferation activity of various cell lines of liver cancer, and showed good therapeutic effect in nude mice model of hepatocellular carcinoma without obvious toxic and side effects; WZ35 inhibited the absorption of glucose in hepatoma cells, and the drug effect glycolysis, phosphorylation and purine metabolism are relatively seriously damaged; WZ35 mainly inhibits YAP from entering the nucleus as a transcription factor activator by activating oxidative stress in liver cancer cells, reducing the transcription of GLUT1, and finally reducing its GLUT1. Tissue microarray and case analysis showed that GLUT1 and YAP were highly expressed and correlated in liver cancer patients, and were associated with poor patient prognosis. The GLUT1-YAP risk model had a high score in predicting prognosis.CONCLUSION:The study confirms that WZ35 is a small molecule glycolysis inhibitor, and through its properties, it mediates metabolic reprogramming dominated by impaired glycolysis, oxidative phosphorylation and purine metabolism to inhibit the proliferation activity of liver cancer cells. Our findings present novel insights into the pathology of liver cancer and potential targets for new therapeutic strategies. GLUT1-YAP has important reference significance for predicting the stages of disease progression in liver cancer patients and have the potential to serve as novel biomarkers for the diagnosis and treatment of liver cancer.

Abstract Background: The significant changes in a series of biochemical activities during the process of tumorigenesis and development, including those in glucose, lipid, and amino acid metabolism can contribute to the ability of the cancer cells to proliferate indefinitely. WZ35 is a new small molecule YAP inhibitor, which can mediate YAP activity to inhibit the growth of gastric cancer, breast cancer and hepatocellular carcinoma. In this article, we explored how WZ35 can modulate YAP activity to influence the metabolism of hepatocellular carcinoma (HCC) cells to inhibit their proliferation activity. Methods: The Gene Expression Omnibus (GEO) data, the Cancer Genome Atlas (TCGA) data, immunohistochemistry (IHC) and preclinical mouse model was utilized to detect the differential expression and vital role of YAP in HCC cells. A series of in vitro and in vivo experiments were performed to explore the antitumor activity of WZ35 and how it can target YAP activity to inhibit the tumor progression. UCSC combined JASPAR public databases were used to predicted possible binding sites of TEAD on GLUT1 promoter. Additionally, seahorse energy metabolism experiments and metabolomics analysis were utilized to explore the possible metabolic changes induced by WZ35. The clinical use of YAP and GLUT1 was verified by bioformatics and tissue microarrays of immunocytochemistry. Results: WZ35 can significantly attenuate the proliferation and growth of HCC. In terms of mechanism(s), it was demonstrated that the suppressive effects of WZ35 on YAP were achieved by promoting ROS production and the drug can exert its significant YAP inhibitory capacity to decrease the level of GLUT1, a glucose transporter located on the surface of cytomembrane, thereby resulting in a significant decrease in the ability of cells to take up glucose and thus perturb the metabolism. Moreover, through bioinformatics mining, it was proposed that YAP/GLUT1 has the potential to serve as promising target for HCC therapy. Conclusions: Our findings indicate that potential inhibitory effects of WZ35 were achieved by affecting the ROS-YAP-GLUT1 signal axis to induce the metabolic reprogramming of hepatocellular carcinoma cells. YAP/GLUT1 could serve as an important molecular target for both the diagnosis and treatment of HCC.

Gastric cancer is the fourth most common cancer and the second most frequent cause of cancer death worldwide. Chemotherapy is an important treatment. However, traditional chemotherapy drugs have low bioavailability and targeting ability. Therefore, we developed curcumin-encapsulated micelles for the treatment of gastric cancer and investigated their antitumor efficacy and active mechanism. Gastric cancer cells were treated with different concentrations of curcumin micelles. MTS cell proliferation assays, flow cytometry (FCM), real time cellular analysis (RTCA) and nude mice xenografts were used to evaluate the effects of curcumin micelles on gastric cancer cell growth in vitro and in vivo. Western blotting was performed to analyze the protein levels of the indicated molecules. A Seahorse bioenergetics analyzer was used to investigate alterations in oxygen consumption and the aerobic glycolysis rate. Curcumin micelles significantly inhibited proliferation and colony formation and induced apoptosis in gastric tumor cells compared to the control groups. We further investigated the mechanism of curcumin micelles on gastric tumor cells and demonstrated that curcumin micelles acted on mitochondrial proteins, causing changes in mitochondrial function and affecting mitochondrial bioenergetics. Furthermore, curcumin micelles decreased mitochondrial membrane potential, increased reactive oxygen species (ROS) generation and disrupted redox equilibrium. The nude mouse model verified that curcumin micelle treatment significantly attenuated tumor growth in vivo. Curcumin micelles suppress gastric tumor cell growth in vitro and in vivo. The mechanism may be related to increasing ROS generation, disrupting redox equilibrium and affecting mitochondrial bioenergetics.