ADT-OH Inhibits Malignant Melanoma Metastasis in Mice Via Suppressing CSE/CBS and FAK/Paxillin Signaling Pathway

Hydrogen sulfide(H2S)is widely recognized as the third endogenous gas signaling molecule and may play a key role in cancer biological processes.ADT-OH(5-(4-hydroxyphenyl)-3H-1,2-dithiocyclopentene-3-thione)is one of the most widely used organic donors for the slow release of H2S and considered to be a potential anticancer compound.In this study,we investigated the antimetastatic effects of ADT-OH in highly metastatic melanoma cells.A tail-vein-metastasis model was established by injecting B16F10 and A375 cells into the tail veins of mice,whereas a mouse footpad-injection model was established by injecting B16F10 cells into mouse footpads.We showed that administration of ADT-OH significantly inhibited the migration and invasion of melanoma cells in the three different animal models.We further showed that ADT-OH dose-dependently inhibited the migration and invasion of B16F10,B16F1 and A375 melanoma cells as evaluated by wound healing and Transwell assays in vitro.LC-MS/MS and bioinformatics analyses revealed that ADT-OH treatment inhibited the EMT process in B16F10 and A375 cells by reducing the expression of FAK and the downstream response protein Paxillin.Overexpression of FAK reversed the inhibitory effects of ADT-OH on melanoma cell migration.Moreover,after ADT-OH treatment,melanoma cells showed abnormal expression of the H2S-producing enzymes CSE/CBS and the AKT signaling pathways.In addition,ADT-OH significantly suppressed the proliferation of melanoma cells.Collectively,these results demonstrate that ADT-OH inhibits the EMT process in melanoma cells by suppressing the CSE/CBS and FAK signaling pathways,thereby exerting its antimetastatic activity.ADT-OH may be used as an antimetastatic agent in the future.