Butyrate and Resveratrol synergize to inhibit colorectal cancer cell proliferation and migration by preventing the nuclear translocation of β-catenin

¹ Department of Health Sciences, Università del Piemonte Orientale, Via Paolo Solaroli 17, 28100 Novara (Italy)
² Department of Health Sciences, Università degli Studi “Magna Graecia” di Catanzaro (Italy)

^* Corresponding Author: Ciro Isidoro, MD, DSc, Email: This email address is being protected from spambots. You need JavaScript enabled to view it.

Abstract

Colorectal cancer (CRC) ranks as the second leading cause of cancer-related mortality worldwide. Disease progression is primarily driven by disruptions in the Wnt/β-catenin pathway, alongside alterations in gut microbiota. Accordingly, epidemiological studies support the evidence that a large portion of colorectal cancers can be prevented by modifying one’s lifestyle, particularly through dietary intake of prebiotics, probiotics, postbiotics, and nutraceuticals that reduce intestinal inflammation and promote intestinal cell homeostasis and barrier integrity. In this context, targeting both microbiota and Wnt/β-catenin pathway represents a promising therapeutic approach. Here, we show that butyrate (the most abundant postbiotic produced by gut microbiota) and resveratrol (a nutraceutical counteracting IL-6-mediated inflammation) can reduce the motility and growth of CRC cells. HCT116 colorectal cancer cells were exposed to sodium butyrate (NaB) and trans-resveratrol (RV). IL-6 was included to mimic the inflammatory tumor microenvironment. The anticancer properties of both NaB and RV, individually or in combination, were tested in wound healing scratch assay, 3D-spheroid forming assay, and immunofluorescence double-staining of epithelial to mesenchymal transition markers and β-catenin. Wound healing assays showed that NaB and RV combination achieved significant inhibition compared to individual inhibitory effects. NaB and RV reduced HCT116 cell migration and proliferation by promoting β-catenin retention in the cytoplasm bound to E-cadherin. In 3D spheroid assays, combination of NaB and RV resulted in higher reduction of spheroid growth than the reducing effects of NaB and RV alone. Immunofluorescence revealed increased cytoplasmic retention of β-catenin bound to E-cadherin and decreased nuclear β-catenin activity, confirmed by downregulation of SNAIL and Cyclin D1. These findings demonstrate a synergistic effect of NaB and RV in inhibiting CRC cell proliferation and migration, supporting their potential as adjuvant dietary interventions in CRC management. These preliminary in vitro data consistently demonstrate the synergism between butyrate and resveratrol in inhibiting CRC cell proliferation and migration, supporting the view that postbiotics and nutraceuticals have great potential as adjuvant and complementary therapeutic agents in CRC treatment.