1273 Crispr/cas9 mediated knockout of ripk4 impairs spheroid formation and wnt/β-catenin signaling in melanoma cells

Many studies indicate the involvement of WNT signaling in oncogenesis and melanoma drug resistance. Receptor-interacting protein kinase (RIPK4), which targets multiple pathways such as Wnt/β-catenin, NFKB MAPK, is might be a alternative target. This supported by fact that RIPK4 levels are elevated cells compared to normal melanocytes. Since role this regulation pathway remain unknown, we tested impact genetic manipulation on Wnt human malignant cells: A375 WM266.4. Our study showed CRISPR/Cas9 mediated knockout (KO) significantly reduces expression phospho-GSK2β, Axin1 β-catenin. In addition, observed impairs Wnt3a-induced activation LPR5/6 β-catenin, was manifested decrease β-catenin Top-Flash reporter system under knockout, control for both WM266.4 cells. Additionally, reduced spheroid formation accompanied ATP increased glucose uptake RIPK4.KO results extend previously published data potential importance melanoma.