A living cell-based fluorescent reporter for high-throughput screening of anti-tumor drugs

Area of Science:

• Biochemistry
• Molecular Biology
• Cancer Research

Background:

• Cellular O-linked β-N-acetylglucosaminylation (O-GlcNAcylation) plays a role in regulating cancer cell proliferation and migration.
• Targeting O-GlcNAcylation presents a potential therapeutic strategy for cancer treatment.

Purpose of the Study:

• To develop a cell-based fluorescent reporter system for sensing cellular O-GlcNAcylation levels.
• To utilize this reporter system for screening natural products as potential anti-cancer agents.

Main Methods:

• Designed a fluorescent reporter using green fluorescent protein (GFP), red fluorescent protein, and neuronal differentiation 1, linked to insulin gene transcription regulation.
• Validated the reporter's ability to detect reduced O-GlcNAcylation in various cell lines.
• Screened 120 natural products using the developed reporter system.

Main Results:

• The fluorescent reporter system successfully sensed changes in cellular O-GlcNAcylation.
• Sesamin was identified as a compound that significantly inhibits O-GlcNAcylation in HeLa and human colorectal carcinoma-116 cells.
• Sesamin demonstrated the ability to repress cancer cell migration in vitro.

Conclusions:

• Developed a novel fluorescent reporter for monitoring O-GlcNAcylation, applicable to gene transcription studies.
• Identified sesamin as a potential anti-cancer compound through a high-throughput drug screening strategy.
• The study provides a new approach for anti-tumor drug discovery and O-GlcNAcylation research.