Assessment of cytotoxic and genotoxic effects of glyphosate-based herbicide on glioblastoma cell lines: Role of p53 in cellular response and network analysis

Toxicology. 2024 Nov:508:153902. doi: 10.1016/j.tox.2024.153902. Epub 2024 Jul 31.

Abstract

Glyphosate, the world's most widely used herbicide, has a low toxicity rating despite substantial evidence of adverse health effects. Furthermore, glyphosate-based formulations (GBFs) contain several other chemicals, some of which are known to be harmful. Additionally, chronic, and acute exposure to GBFs among rural workers may lead to health impairments, such as neurodegenerative diseases and cancer. P53 is known as a tumor suppressor protein, acting as a key regulator of the cellular response to stress and DNA damage. Therefore, mutations in the TP53 gene, which encodes p53, are common genetic alterations found in various types of cancer. Therefore, this study aimed to evaluate the cytotoxicity and genotoxicity of GBF in two glioblastoma cell lines: U87MG (TP53-proficient) and U251MG (TP53-mutant). Additionally, the study aimed to identify the main proteins involved in the response to GBF exposure using Systems Biology in a network containing p53 and another network without p53. The MTT assay was used to study the toxicity of GBF in the cell lines, the clonogenic assay was used to investigate cell survival, and the Comet Assay was used for genotoxicity evaluation. For data analysis, bioinformatics tools such as String 12.0 and Stitch 5.0 were applied, serving as a basis for designing binary networks in the Cytoscape 3.10.1 program. From the in vitro test analyses, it was observed a decrease in cell viability at doses starting from 10 ppm. Comet Assay at concentrations of 10 ppm and 30 ppm for the U251MG and U87MG cell lines, respectively observed DNA damage. The network generated with systems biology showed that the presence of p53 is important for the regulation of biological processes involved in genetic stability and neurotoxicity, processes that did not appear in the TP53-mutant network.

Keywords: Comet assay; Glyphosate; System biology; TP53; in silico; in vitro.

MeSH terms

  • Cell Line, Tumor
  • Cell Survival* / drug effects
  • Comet Assay
  • DNA Damage* / drug effects
  • Dose-Response Relationship, Drug
  • Glioblastoma* / genetics
  • Glioblastoma* / pathology
  • Glycine* / analogs & derivatives
  • Glycine* / toxicity
  • Glyphosate*
  • Herbicides* / toxicity
  • Humans
  • Mutation
  • Tumor Suppressor Protein p53* / genetics
  • Tumor Suppressor Protein p53* / metabolism

Substances

  • Glycine
  • Herbicides
  • Glyphosate
  • Tumor Suppressor Protein p53
  • TP53 protein, human