Caffeine enhances migration, invasion, and tube formation by upregulating MMP-2 in EVT-like HTR-8/SVneo cells

Abstract

Caffeine is consumed widely during pregnancy, yet its effects on cellular processes relevant to early placental development, particularly on extravillous trophoblast (EVT) function, remain incompletely understood. This study examined the impact of caffeine on EVT-like HTR-8/SVneo cells, focusing on key cellular processes including migration, invasion, and matrix-interactive behaviors relevant to trophoblast function. Treatment with 1 mM caffeine enhanced cell motility and tube formation in EVT-like cells without affecting cell viability or inducing apoptosis. These functional changes were associated with a selective upregulation of MMP-2 expression and activity, while MMP-9 remained unchanged. Pharmacological inhibition of MMP-2 abolished caffeine-induced increases in migration, invasion, and tube formation, supporting a functional role for MMP-2 in these responses. At the signaling level, caffeine-induced MMP-2 upregulation was attenuated by inhibition of PI3K/Akt signaling, with a minor contribution from PKA, whereas MEK1/2 inhibition had minimal effect, supporting the involvement of distinct signaling pathway in EVT-like cells. The observed trophoblast-specific response contrasts with the inhibitory effects of caffeine reported in cancer and endothelial cells. Collectively, these findings identify a previously uncharacterized mechanism by which caffeine modulates signaling and matrix-remodeling−associated behaviors in EVT-like cells in vitro.