BHMPS Inhibits Breast Cancer Migration and Invasion by Disrupting Rab27a-Mediated EGFR and Fibronectin Secretion

SIMPLE SUMMARY: Numerous studies targeting Rab GTPases and its multiple effectors have been attempted since exocytosis has been shown to alter tumor malignancy by modulating cancer cell behavior and tumor microenvironment. Here, we demonstrated that BHMPS inhibits migration and invasion of breast cancer cells by blocking the interaction between Rab27a and Slp4. BHMPS interfered with vesicle trafficking and secretion by decreasing FAK and JNK activation. In addition, BHMPS suppressed tumor growth in Rab27a-overexpressing MDA-MB-231 xenograft mice. This study highlighted the importance of understanding the mechanisms of Rab27a-mediated metastasis in improving the therapeutic options for metastatic cancers. ABSTRACT: Our previous work demonstrated that (E)-N-benzyl-6-(2-(3, 4-dihydroxybenzylidene) hydrazinyl)-N-methylpyridine-3-sulfonamide (BHMPS), a novel synthetic inhibitor of Rab27aSlp(s) interaction, suppresses tumor cell invasion and metastasis. Here, we aimed to further investigate the mechanisms of action and biological significance of BHMPS. BHMPS decreased the expression of epithelial-mesenchymal transition transcription factors through inhibition of focal adhesion kinase and c-Jun N-terminal kinase activation, thereby reducing the migration and invasion of breast cancer. Additionally, knockdown of Rab27a inhibited tumor migration, with changes in related signaling molecules, whereas overexpression of Rab27a reversed this phenomenon. BHMPS effectively prevented the interaction of Rab27a and its effector Slp4, which was verified by co-localization, immunoprecipitation, and in situ proximity ligation assays. BHMPS decreased the secretion of epidermal growth factor receptor and fibronectin by interfering with vesicle trafficking, as indicated by increased perinuclear accumulation of CD63-positive vesicles. Moreover, administration of BHMPS suppressed tumor growth in Rab27a-overexpressing MDA-MB-231 xenograft mice. These findings suggest that BHMPS may be a promising candidate for attenuating tumor migration and invasion by blocking Rab27a-mediated exocytosis.