Mesalazine and Lactoferrin as Potential Adjuvant Therapy in Colorectal Cancer: Effects on Cell Viability and Wnt/β-Catenin Pathway

Abstract

Colorectal cancer (CRC) remains one of the leading causes of cancer-related deaths, meaning it is essential to explore all possible strategies for its prevention and treatment. Unfortunately, risk factors such as an unhealthy lifestyle, lack of exercise, and obesity-which are increasingly prevalent in developed countries-contribute to CRC development. The aim of this study was to evaluate the effect of a mesalazine (MES) and lactoferrin (LACT) combination on the viability of CRC cells and healthy intestinal epithelial cells, as well as to assess the expression profile of target genes within the Wnt/β-catenin pathway. Additionally, this study aimed to preliminarily analyze the mechanism of action underlying the combined effects of these compounds. In this study, we used three CRC cell lines (HCT-116, DLD-1, and HT-29) along with the healthy intestinal epithelial cell line CCD 841 CoN. These cells were treated with MES and LACT separately, as well as in combination. We demonstrated that the combination of MES and LACT reduced the viability of CRC cells more effectively than either compound alone, while slightly increasing the viability of normal intestinal epithelial cells. The synergistic effect of MES and LACT may serve as a foundation for developing new treatment strategies for CRC, utilizing compounds with a high safety profile.

Keywords: colorectal cancer; iron metabolism; lactoferrin; mesalazine.

Figure 1

CRC treatment options and risk factors. The figure was partly generated using Servier Medical Art, provided by Servier, licensed under a Creative Commons Attribution 3.0 unported license.

Figure 2

Changes in cell viability of CRC cell lines HT-29, DLD-1, and HCT-116, as well as the normal epithelial cell line CCD 841 CoN, after 24 h of exposure to MES, measured by the MTT assay. 5-FU was used as a positive control for cytotoxicity. Each experimental condition was performed in three biological replicates.

Figure 3

Changes in cell viability of CRC cell lines (a) HT-29, (b) DLD-1, and (c) HCT-116 and the normal epithelial cell line (d) CCD 841 CoN after 24 h of exposure to MES (30 mM), LACT (400 µg/mL), and their combination (MES with LACT), measured by the MTT assay. Data are presented as mean ± SD from three independent experiments. Although statistical analyses were performed, significance markers were not added to the figure to preserve clarity and avoid confusion regarding multiple comparisons.

Figure 4

Changes in mRNA levels of (a) CCND1 and (b) MYC in the CCD 841 CoN cell line, (c) CCND1 and (d) MYC in the DLD-1 cell line, (e) CCND1 and (f) MYC in the HCT-116 cell line, and (g) CCND1 and (h) MYC in the HT-29 cell line after 24 h of treatment with 30 mM MES, 400 µg/mL LACT, 30 mM MES + 400 µg/mL LACT, and non-treated control cells (CON). Gene expression levels were normalized to the reference gene TBP and calculated using the 2^−ΔΔCT method. Box and whisker plots display the median, lower and upper quartiles, and minimum and maximum values. Abbreviations: MES—mesalazine; LACT—lactoferrin; CON—control (untreated).

Figure 5

Graphical representation of MES interaction with LACT (a), CCND1 (b), and MYC (c). Red arrow (a) indicates a ligand (MES); green color indicates hydrogen bonds.