Royal jelly with ellagic acid inhibits the glycolytic pathway and induces apoptosis through multiple pathways in colorectal cancer.

Abstract

Background: Developing novel chemotherapeutics with high anticancer efficacy and low toxicity remains a critical challenge in oncology. Natural products have shown promise due to their multitargeted activity and favorable safety profiles. The present study investigates the combined anticancer effects of royal jelly (RJ) and ellagic acid (EA), two potent antioxidants of animal and plant origin.

Materials and methods: Royal jelly (RJ) and ellagic acid (EA) were applied to HT29 (ATCC HTB-38, Human colorectal adenocarcinoma), HCT116 (ATCC CCL-247, Human colorectal carcinoma) and BEAS-2B (ATCC CRL-3588, human bronchial epithelium) cell lines, and their antiproliferative effects were evaluated using a xCELLigence Real-Time Cell Analyzer (RTCA MP). The effect of the combination of RJ and EA on the glycolytic pathway was determined using a Seahorse XFe24 Analyzer, and the apoptotic process was evaluated by DNA laddering and the expression of the Bcl-2 and Bax genes in the apoptotic pathway through real-time quantitative PCR (RT-qPCR). The transcriptome profiling of the combination of RJ and EA on colorectal cancer cells was performed by Total RNA Sequencing analysis.

Results: RJ with EA, when used in combination, significantly reduced the extracellular acidification rate (ECAR), effectively inhibiting aerobic glycolysis, especially in HCT116, and induced apoptosis in HCT116 and HT29 cells by increasing the Bax/Bcl-2 ratio compared to cases treated with EA or RJ alone (p < 0.05). GSEA analyses revealed that the treatment of both cell lines increased the expression of apoptosis and p53 pathway-related genes while suppressing the genes associated with the E2F target, G2M checkpoint, oxidative phosphorylation, and MYC target mechanism, indicating a directly proportional relationship with the antiproliferative effect on cancer cells and increased apoptosis.

Conclusion: RJ with EA used in combination demonstrates potent anticancer effects in colorectal cancer by suppressing glycolysis and activating apoptosis, with apparent therapeutic potential as a novel cancer treatment strategy.