Abstract

A Robust High Throughput Image Based Screening Assay to Identify Inhibitors of Gastric Cancer Cell Lines

Sravanthy Manesh, Research Scientist, Experimental Therapeutics Centre

Image-based assays are rapidly finding its niche in early stage drug discovery screening campaigns. The combination of one or more fluorescent probes, an automated imager and image analysis algorithms provides a robust and competent platform to unravel the effect of test compounds on cytological events and signaling pathways. Compounds that cause perturbations in actin polymerization leading to apoptosis are known to be attractive targets for cancer therapy. The assay described here is a robust image based assay to identify inhibitors of actin polymerization. 24 different Gastric cancer cell lines were treated with known anti-cancer drugs in a dose dependent assay. The cells were then fixed followed by staining with fluorescent-tagged phalloidin probe and nuclei stain To-Pro-3. The cells were then imaged using GE Healthcare IN Cell 2000. The nuclei count and actin intensity were quantified using GE IN Cell Developer software. The image-based assay described here is a highly sensitive, reproducible and reliable secondary assay that can be used for primary screening hit validation. This presentation will showcase the inhibitors identified using this image-based assay.