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

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.