Therapeutic potential of combined treatment with chlorogenic acid and 5-fluorouracil in colorectal cancer through Wnt/β-catenin pathway modulation

Abstract

Background

Colorectal cancer (CRC) has become a significant public health concern due to its rising incidence and mortality rates. Given the limitations of chemotherapy, exploring novel therapeutic strategies is essential. The Wnt/β-catenin pathway is often deregulated in CRC, contributing to its development and progression. Chlorogenic acid has demonstrated a promising modulatory effect on this pathway.

Purpose

In this context, this study evaluated the effect of chlorogenic acid in combination with the chemotherapeutic 5-fluorouracil on cell viability and proliferation, as well as on the modulation of the Wnt/β-catenin pathway.

Study design

Combined treatments were performed on CRC cell lines SW480 and HT-29. Both lines exhibit alterations in the APC gene, a key component of the Wnt/β-catenin pathway; however, SW480 displays higher transcriptional activity of the pathway.

Methods

Cellular and molecular biology techniques were employed in this study. The MTT assay was used to assess cell viability, while flow cytometry was used to analyze cell cycle progression as a measure of cellular proliferation. qRT-PCR was conducted to quantify the expression of genes involved in the Wnt/β-catenin signaling pathway and its target genes. Western blotting was performed to measure β-catenin protein levels, and fluorescence microscopy was applied to evaluate the fluorescence intensity and cellular localization of β-catenin.

Results

The results showed that co-treatment decreased cell viability and altered the cycle in both lines, with SW480 cells showing greater sensitivity. Furthermore, differential modulation of pathway-related gene expression levels was observed, including decreased β-catenin and TCF7L2. Changes in the expression of genes associated with drug resistance and tumor metabolism were also identified, significant decreases of ABCG2 in HT-29 and of SLC16A1 in SW480.

Conclusion

The results suggest that chlorogenic acid enhances the efficacy of conventional therapy by modulating the Wnt/β-catenin pathway, which could potentially impact the carcinogenesis of CRC.