Carmofur inhibits ovarian cancer cell proliferation, migration, and invasion via disrupting the YAP-1/TEAD pathway: in vitro and in silico approaches

Abstract

The current study aims to present a novel approach to alleviate the aggressiveness of ovarian cancer by pharmacological disruption of the Yes-Associated Protein (YAP) - 1/Transcriptional Enhancer Factor domain (TEAD) heterodimer structure using Carmofur. The Ovcar-3 cell line was incubated with progressively diluted Carmofur solutions. Proliferation inhibition was assessed using the MTT assay. Flow cytometry was employed to determine the pro-apoptotic properties. Cellular morphological alterations were assessed with confocal microscopy. The migration inhibition status was assessed via the scratch assay, whilst the colony formation inhibition level was evaluated using the colony assay. The western blotting technique was employed to determine YAP-1 expression levels. The disruptive impact of Ceramide and Carmofur on the YAP-1/TEAD heterodimer structure was elucidated via molecular modeling, considering three interfaces: the beta-strand, the alpha-helix, and the Omega-loop. Furthermore, validation was conducted by assessing the disruptive impact of Verteporfin on the YAP-1/TEAD heterodimer. The findings indicated that Carmofur diminished cell viability in a concentration- and time-dependent manner, with IC50 values of 17.26 & micro;M at 24 h, 10.15 & micro;M at 48 h, and 9.22 & micro;M at 72 h, respectively. Carmofur showed a 26.65% apoptotic effect at the IC50 value over 24 h. Additionally, Carmofur diminished the relative YAP-1 levels from 9.72 to 2.98 in the Ovcar-3 cell line and exhibited an anti-metastatic effect by suppressing colony formation and migratory capabilities. This is the first study to provide in silico and in vitro evidence that Carmofur induces apoptosis by disrupting the YAP-1/TEAD heterodimer, thereby inhibiting the proliferation and migration of Ovcar-3 cells.