In vitro cytogenetic effects of extract from Raphanus sativus roottuber

In this study, the cytogenotoxic and mutagenic potential of the aqueous extract (RSÖ) obtained from underground hypertrophic roots of Raphanus sativus (radish) was investigated under in vitro conditions. The experiments were conducted using auxotrophic mutant strains of Salmonella typhimurium (TA98 and TA100), as well as human breast cancer (MCF7) and mouse embryonic fibroblast (NIH3T3) cell lines, which were exposed to subtoxic concentrations of RSÖ for brief periods. Cell viability was determined using a CCK-8 assay, while genotoxicity was evaluated using Ames, comet, and micronucleus (MN) assays. The application of RSÖ at concentrations ranging from 0.5 to 2 mg/ml for 24 or 48 hours did not result in significant levels of cytotoxicity in either the MCF7 or NIH3T3 cell lines. Notably, RSÖ was observed to enhance the viability of MCF7 cells at low doses. Furthermore, RSÖ demonstrated a protective effect against hydrogen peroxide (H2O2)-induced cell death, with a significant increase in cell viability (p≤0.001). However, in the Salmonella test, RSÖ did not demonstrate mutagenic potential in the TA98 strain, yet it significantly elevated the number of revertant colonies in the TA100 strain, indicative of potential mutagenicity. In antimutagenicity assays, it was ascertained that RSÖ notably diminished the mutagenic activity in the TA98 strain at a high dose (4 mg/petri) (p≤0.001). In the comet assay, it was observed that high concentrations of RSÖ caused genotoxicity in MCF7 and NIH3T3 cells. However, when combined with H2O2, the extract did not reduce oxidative stress-induced DNA damage. In the MN assay, low concentrations (0.5-1 mg/ml) of RSÖ induced an antigenotoxic response by reducing micronucleus formation in MCF7 cells, while high concentrations (2 mg/ml) increased cell proliferation. However, in NIH3T3 cells, 48-hour treatment with 1-2 mg/ml RSÖ resulted in a substantial increase in MN and micronucleated binucleated cell (MNBC) frequencies, accompanied by a significant induction of cellular toxicity and genotoxicity (p≤0.01). These data reveal the dose-dependent genotoxicity and antigenotoxicity dynamics of RSÖ, demonstrating that while it exhibits a DNA-protective effect at low concentrations, it poses a genotoxic risk at high doses with cell type specificity and that this dual behavior necessitates strict parametric optimization of the therapeutic window.