Integrated Metabolomics and Proteomics Analyses to Reveal Anticancer Mechanism of Hemp Oil Extract in Colorectal Cancer

Abstract Background Cannabis sativa L. has a lengthy medical history in Chinese folk medicine. Industrial cannabis, also known as hemp, refers to cannabis strains with no addictive effect and holds great economic and medical value. Hemp exhibited multiple pharmaceutical activities with anti-inflammatory, antioxidant, and anti-tumor effects. Hemp oil extract (HOE), a mixture of cannabinoid-rich components extracted from hemp, has shown potential in treating colorectal cancer (CRC). However, the specific anticancer mechanisms of HOE remain unclear. This research aims to elucidate the anticancer mechanisms of HOE in the treatment of CRC by employing an integrated omic approach, analyzing changes in cellular metabolites and proteins. Methods In this study, we employed mass spectrometry-based omic approaches, specifically metabolomics and proteomics, to comprehensively investigate the global effects of HOE on colorectal cancer cells. Bioinformatics analysis including bulk RNA-seq and single-cell RNA-seq were utilized to unravel specific gene expression differences and heterogeneity in CRC. The inferred conclusions were confirmed by utilizing flow cytometry, western blot, and immunohistochemistry techniques. Results The in vitro and in vivo experiments indicated that HOE induced significant changes in purine metabolism pathways, down-regulated c-MYC and inhibited the expression of cell cycle-related proteins, including CCND1, CDK4, CDK6, which herein arrested cell cycle in G1 phase. Conclusion Through Integrating Metabolomics and Proteomics approaches, a comprehensive analysis of anticancer mechanisms of HOE on CRC was conducted. The study demonstrated that HOE blocks the cell cycle in the G1 phase by inhibiting c-MYC, leading to the inhibition of colorectal cancer cell proliferation. These findings provide experimental evidence for the potential use of hemp in medicine.