PO-415 New anti-migratory and anti-invasive effects of a fascin inhibitor on colorrectal cancer cells

Introduction Serrated adenocarcinoma (SAC) is a histological subtype of colorrectal carcinoma characterised by its poor prognosis, prominent invasive front and over-expression of fascin, the key protein involved in actine bundling needed for cell migration and invasion. Given that the frequency of KRAS or BRAF mutations in SAC are higher than in conventional colorectal carcinoma, this tumour type is usually resistant to anti-EGFR therapy. For these reason, anti-fascin treatment could be an interesting aproach to treat SAC and other tumours over-expressing fascin. In this work we have charaterized the anti-migratory and anti-invasive properties of compound G2 (N-(1-(4-(trifluoromethyl)benzyl)−1H-indazol-3-yl)furan-2-carboxamide), a patented anti-fascin molecule and compared its effect with migrastatine, a typical fascin inhibitor. Material and methods We have used molecular modelling to predict the fascin aminoacids involved in G2 binding. A qPCR assay was carried to find out which from eigth different colorectal cancer cell lines expressed the highest amount of fascin. After cell viability assay, scratch and IBIDI assays were performed to evaluate the anti-migratory effect of the compounds. Immunofluoresce for fascin was used for assessing lamelipodia formation. Anti-invasive effect was evaluated using cell invasion assay Transwell with matrigel and a myoma organotypic invasion model. Results and discussions Molecular modelling using blind docking calculations identified a region in fascin possibly involved in G2 binding. HCT116 cells expressed the highest fascin levels and its migration capacity was clearly reduced by migrastatin and G2, the latter even at lower concentrations. Still, G2 inhibited the migration of all the cell lines demonstrating that G2 affected fascin functional capacity. Invasion and confocal studies were performed with HCT116 cells and both inhibitors strongly abolished the protrusion of lamellipodium (p=0.003). The Transwell Matrigel invasion assay also evidenced the anti-invasive effects of migrastatin and G2. Myoma discs showed that both compounds were similarly able to significantly decrease both the invasion depth and invasion area of HCT116 cells at 50% (p Conclusion This study demonstrates an interesting anti-migratory and anit-invasive effect of G2 in a similar extent to migrastatin and provides significant evidence that G2 is an interesting candidate for further investigation/chemical modifications to develop new fascin-specific therapies for colorectal cancer.