Development of human cell biosensor system for genotoxicity detection based on DNA damage-induced gene expression

Area of Science:

• Biotechnology
• Toxicology
• Molecular Biology

Background:

• Environmental genotoxic agents pose significant health risks.
• Rapid assessment of chemical genotoxicity is crucial for drug development and environmental monitoring.
• The p21 tumor suppressor gene is a key indicator of DNA damage and cell cycle arrest.

Purpose of the Study:

• To develop a human cell-based biosensor for simple and rapid detection of genotoxic agents.
• To utilize the p21 promoter's response to DNA damage for biosensing.
• To establish a reliable system for assessing chemical genotoxicity.

Main Methods:

• Transfection of HepG2 cells with a p21-EGFP reporter construct (p21HepG2GFP).
• Induction of DNA damage using known genotoxic agents (MMS, BaP, cisplatin, vinblastine).
• Measurement of EGFP fluorescence intensity and cell viability (MTS assay).

Main Results:

• Significant EGFP increase observed with methylmethane sulphonate (MMS), benzo(a)pyrene (BaP), and cisplatin.
• Dose-dependent responses noted for BaP and cisplatin.
• Vinblastine showed minimal EGFP induction, with higher concentrations reducing cell viability.

Conclusions:

• The p21HepG2GFP cell line serves as a robust biosensor for detecting genotoxicity.
• This assay provides a fast and simple method for identifying genetic damage induced by chemical agents.
• The system is valuable for environmental monitoring and chemical safety assessments.