[Development of fluorescence imaging based assay for screening compounds with anti-migration activity]

Area of Science:

• Biochemistry
• Cell Biology
• Pharmacology

Context:

• Tumor cell migration is a critical process in cancer metastasis.
• Developing efficient screening methods is crucial for identifying anti-cancer drugs.
• High-throughput screening (HTS) enables rapid evaluation of numerous compounds.

Purpose:

• To develop and validate a fluorescent imaging-based high-throughput screening (HTS) method for identifying anti-migratory compounds.
• To optimize key parameters of the screening assay, including incubation and migration times.
• To assess the inhibitory effect of paclitaxel on tumor cell migration and screen natural product components.

Summary:

• A novel fluorescent imaging HTS method was established using 96-well Transwell plates to detect compounds inhibiting cell migration.
• The method's correlation, precision, and stability were evaluated, and parameters like Hoechst 33342 incubation and migration times were optimized.
• The anti-migratory activity of paclitaxel was confirmed with an IC50 of 0.717 µmol/L. Screening 24 Rhizoma Alismatis components identified one with anti-migratory potential.

Impact:

• The developed HTS method offers a precise, stable, and potentially linear assay for evaluating anticancer compounds targeting cell migration.
• This approach facilitates the discovery of novel anti-metastatic agents from natural products and synthetic libraries.
• The validated method can accelerate the drug discovery pipeline for anti-cancer therapies.