# Toxic effects of Helix aspersa snail egg hydrolyzates obtained by static in vitro digestion on Caco-2 colorectal adenocarcinoma cells

## TL;DR

This study shows that hydrolyzed snail egg extracts may selectively harm colon cancer cells without affecting normal cells.

## Contribution

The novel contribution is demonstrating the anticancer potential of snail egg hydrolyzates against Caco-2 cells.

## Key findings

- Hydrolyzates reduced plasma membrane integrity in Caco-2 cells but not in normal IEC-6 cells.
- Hydrolyzates altered apoptotic protein concentrations and mitochondrial membrane potential in cancer cells.
- Bioactive compounds released during digestion likely caused the observed anticancer effects.

## Abstract

Colorectal cancer is the third most common cancer. Anticancer compounds obtained after the digestion of snail eggs may hurt the growth and development of colon cancer cells. The research aimed to prepare hydrolyzates from Helix aspersa maxima and Helix aspersa aspersa snail eggs using static in vitro simulation of gastrointestinal digestion. Redox status indicators, concentration of total protein, total carbohydrates, uronic acids, allantoin, and glycolic acid, and electrophoretic profile of proteins and glycoproteins were compared in hydrolyzates and non-digested extracts. The effect of hydrolyzates on the integrity of the plasma membrane of Caco-2 human epithelial colorectal adenocarcinoma cells and the monolayer of IEC-6 rat intestinal cells was determined. The influence of hydrolyzates on the production of reactive oxygen species (ROS) by Caco-2 cells, the potential of their mitochondrial membrane, and the production of apoptotic proteins were examined. Incubation with hydrolyzates for 72 h reduced the plasma membrane integrity of Caco-2 cells but not normal cells, did not affect ROS content, decreased mitochondrial membrane potential (the tendency for H. a. aspersa hydrolyzate), and altered the concentration of some apoptotic proteins of intrinsic and extrinsic apoptosis pathways. Bioactive compounds released during digestion and their interactions may have been responsible for anticancer effects.

The online version contains supplementary material available at 10.1038/s41598-025-11605-7.