Low-density lipoprotein receptor-related protein 6 is a novel coreceptor of protease-activated receptor-2 in the dynamics of cancer-associated β-catenin stabilization

Abstract

Protease-activated receptor-2 (PAR₂) plays a central role in cancer; however, the molecular machinery of PAR₂-instigated tumors remains to be elucidated. We show that PAR₂ is a potent inducer of β-catenin stabilization, a core process in cancer biology, leading to its transcriptional activity. Novel association of low-density lipoprotein-related protein 6 (LRP6), a known coreceptor of Frizzleds (Fz), with PAR₂ takes place following PAR₂ activation. The association between PAR₂ and LRP6 was demonstrated employing co-immunoprecipitation, bioluminescence resonance energy transfer (BRET), and confocal microscopy analysis. The association was further supported by ZDOCK protein-protein server. PAR₂-LRP6 interaction promotes rapid phosphorylation of LRP6, which results in the recruitment of Axin. Confocal microscopy of PAR₂-driven mammary gland tumors in vivo, as well as in vitro confirms the association between PAR₂ and LRP6. Indeed, shRNA silencing of LRP6 potently inhibits PAR₂-induced β-catenin stabilization, demonstrating its critical role in the induced path. We have previously shown a novel link between protease-activated receptor-1 (PAR₁) and β-catenin stabilization, both in a transgenic (tg) mouse model with overexpression of human PAR₁ (hPar1) in the mammary glands, and in cancer epithelial cell lines. Unlike in PAR₁-G_α13 axis, both G_α12 and G_α13 are equally involved in PAR₂-induced β-catenin stabilization. Disheveled (DVL) is translocated to the cell nucleus through the DVL-PDZ domain. Collectively, our data demonstrate a novel PAR₂-LRP6-Axin interaction as a key axis of PAR₂-induced β-catenin stabilization in cancer. This newly described axis enhances our understanding of cancer biology, and opens new avenues for future development of anti-cancer therapies.