The PMA Phorbol Ester Tumor Promoter Increases Canonical Wnt Signaling Via Macropinocytosis

Abstract

Activation of the Wnt pathway lies at the core of many human cancers. Interestingly, Wnt, cell adhesion, and macropinocytosis are often active in the same processes, and understanding how Wnt signaling and membrane trafficking cooperate should improve our understanding of embryonic development and cancer. Here we show that a macropinocytosis activator, the tumor promoter Phorbol 12-myristate 13-acetate (PMA), enhances Wnt signaling. Experiments using the Xenopus embryo as an in vivo model showed marked cooperation between the PMA phorbol ester and Wnt signaling, which was blocked by inhibitors of macropinocytosis, Rac1 activity, and lysosome acidification. The crosstalk between canonical Wnt, the Protein Kinase C (PKC) pathway, focal adhesions, lysosomes, and macropinocytosis suggests possible therapeutic targets for cancer progression in Wnt-driven cancers.

Addition of exogenous diacylglycerol enhances Wnt/β-catenin signaling through stimulation of macropinocytosis

“… 5 , 6 , 7 In a separate study, we provide evidence that PMA increases Wnt signaling, which can be blocked by inhibitors of macropinocytosis, MVB formation, membrane trafficking, and lysosome acidification. 40 Here, we showed that addition of DAG strongly potentiates macropinocytosis and Wnt signaling. These findings have potential implications for the prevention of cancer progression.…”

“…These effects were confirmed by western blot ( Figure 4 G). In cells transfected with myristoylated-GFP, 40 which marks the plasma membrane, DAG addition strongly increased macropinosome plasma membrane activity within minutes ( Figure 4 , compare panels H and I; Video S1 ). The increase in macropinosome number on the plasma membrane is observed almost immediately after addition of DAG ( Video S2 ).…”