Cracking Cancer with CRK

Studies have shown that Crk is overexpressed in many breast cancers and is responsible for several key signalling events. As such, the effects of Crk knockdown in breast tumour cells warrants further analysis. This project studied the outcomes of Crk knockdown in MDA-MB-231 human breast adenocarcinoma cell line on cellular adhesion, morphology and intracellular signalling. This was accomplished by establishing cultured cell groups which consisted of tumour cells infected with vesicular stomatitis viruses (VSV) containing a small hairpin RNA (shRNA) against Crk (knockdown), and a control group infected with the empty pSup vector. The cell groups were compared in terms of their efficiency in matrix-binding in adhesion assays on collagen, fibronectin and glass in time intervals of 20, 40 and 60 minutes in duplicates a number of times. Later, immunofluorescence staining, assessed by confocal microscopy, was carried out to identify changes in cellular morphology and immunoblotting was performed to monitor levels of phosphorylated-TYR in the cell groups. In conclusion, the shRNA- Crk resulted in the significant reduction of cell adhesion and migration and lead to dramatic cytoskeletal rearrangements. Variations in levels of Cas and FAK protein binding interactions were also identified. Further, understanding these protein interactions will help us derive safer anti-cancer treatments and identify potential anti-metastatic therapeutic targets.