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Updated: Jun 3, 2026
Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
A cell-based high-throughput assay for the screening of small-molecule inhibitors of p53-MDM2 interaction
Jing Li1, Shuyong Zhang, Linghuan Gao
1Stake Key Laboratory of Drug Research, the National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
Abstract:
The p53 tumor suppressor is a potent transcription factor that regulates cell growth inhibition and apoptosis. The oncoprotein MDM2 suppresses p53 activity by direct inhibition of its transcriptional activity and enhances the degradation of p53 via the ubiquitin-proteosome pathway. Overexpression of MDM2, found in many human tumors, impairs p53-mediated cell death effectively. Inhibition of the p53-MDM2 interaction can stabilize p53 and may offer a novel strategy for cancer therapy. To search for new inhibitors of the p53-MDM2 interaction, the authors developed a cell-based high-throughput assay system based on mammalian two-hybrid technology. They also used a dual-luciferase reporter system to rule out false- positive hits due to the cytotoxic effect of compounds. Using this assay, they screened a library consisting of 3840 compounds and identified one compound that activates p53 pathway and induces growth arrest in tumor cells.
Insights
Researchers screened compounds to find inhibitors of the p53-MDM2 interaction. They identified a novel compound that activates the p53 pathway, inducing tumor cell growth arrest, offering a potential cancer therapy strategy.
Area of Science:
- Molecular Biology
- Cancer Research
- Drug Discovery
Background:
- The p53 tumor suppressor is crucial for regulating cell growth and apoptosis.
- MDM2 oncoprotein inhibits p53 activity and promotes its degradation, a common issue in human tumors.
- Inhibiting the p53-MDM2 interaction is a promising cancer therapeutic strategy.
Purpose of the Study:
- To develop a high-throughput screening assay for identifying inhibitors of the p53-MDM2 interaction.
- To discover novel compounds that can reactivate the p53 pathway in cancer cells.
Main Methods:
- Developed a cell-based mammalian two-hybrid assay for high-throughput screening.
- Utilized a dual-luciferase reporter system to validate compound efficacy and rule out cytotoxicity.
- Screened a library of 3840 compounds.
Main Results:
- Identified one compound that effectively inhibits the p53-MDM2 interaction.
- The identified compound activates the p53 pathway.
- This compound induced growth arrest in tumor cells.
Conclusions:
- The developed high-throughput assay is effective for identifying p53-MDM2 interaction inhibitors.
- A novel compound was discovered that reactivates p53 and inhibits tumor cell growth.
- This finding supports targeting the p53-MDM2 interaction as a viable cancer treatment approach.
