Integrated multi-omics analysis suggests the involvement of PI3K-Akt/p21 signaling in the anti-colorectal cancer effects of Diaphragma Juglandis Fructus extract.

BACKGROUND

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, underscoring the need for more effective and well-tolerated therapeutic strategies. Diaphragma Juglandis Fructus (DJF), a by-product of walnut processing used in traditional medicine, is rich in polyphenolic compounds, yet its potential effects on CRC have not been fully characterized. This study aimed to evaluate the anti-colorectal cancer activity of the ethanol extract of DJF (EEDJF) and explore the underlying molecular mechanisms using an integrated multi-omics strategy.

METHODS

Network pharmacology, molecular docking, and label-free proteomics were combined with cell-based assays. Functional relevance of AKT signaling was evaluated using siRNA knockdown, MK2206, and SC79.

RESULTS

Fifty-three putative bioactive compounds and 154 CRC-related targets were identified, with AKT1, EGFR, and SRC emerging as candidate hub proteins. EEDJF significantly inhibited proliferation in HCT116 and SW480 colorectal cancer cells, induced apoptosis, and promoted G1/S cell cycle arrest. In contrast, EEDJF showed minimal cytotoxicity toward HaCaT epithelial cells under the tested conditions. Proteomic and enrichment analyses consistently highlighted the PI3K-Akt-related signaling among the most significantly associated pathways. Modulation of AKT activity altered the cellular response to EEDJF, supporting the involvement of this pathway.

CONCLUSION

EEDJF exerts anti-colorectal cancer effects in vitro, potentially associated with regulation of PI3K-Akt/p21 signaling. These findings provide a basis for further studies on the bioactive constituents, pharmacological mechanisms, and in vivo efficacy of DJF-derived preparations.