Inhibition of Wnt/ß-Catenin Signaling in Neuroendocrine Tumors in vitro: Antitumoral Effects.

ABSTRACT

BACKGROUND AND AIMS: Inhibition of Wnt/ß-catenin signaling by specific inhibitors is currently being investigated as an antitumoral strategy for various cancers. The role of Wnt/ß-catenin signaling in neuroendocrine tumors still needs to be further investigated. METHODS: This study investigated the antitumor activity of the porcupine (PORCN) inhibitor WNT974 and the ß-catenin inhibitor PRI-724 in human neuroendocrine tumor (NET) cell lines BON1, QGP-1, and NCI-H727 in vitro. NET cells were treated with WNT974, PRI-724, or small interfering ribonucleic acids against ß-catenin, and subsequent analyses included cell viability assays, flow cytometric cell cycle analysis, caspase3/7 assays and Western blot analysis. RESULTS: Treatment of NET cells with WNT974 significantly reduced NET cell viability in a dose- and time-dependent manner by inducing NET cell cycle arrest at the G1 and G2/M phases without inducing apoptosis. WNT974 primarily blocked Wnt/ß-catenin signaling by the dose- and time-dependent downregulation of low-density lipoprotein receptor-related protein 6 (LRP6) phosphorylation and non-phosphorylated ß-catenin and total ß-catenin, as well as the genes targeting the latter (c-Myc and cyclinD1). Furthermore, the WNT974-induced reduction of NET cell viability occurred through the inhibition of GSK-3-dependent or independent signaling (including pAKT/mTOR, pEGFR and pIGFR signaling). Similarly, treatment of NET cells with the ß-catenin inhibitor PRI-724 caused significant growth inhibition, while the knockdown of ß-catenin expression by siRNA reduced NET tumor cell viability of BON1 cells but not of NCI-H727 cells. CONCLUSIONS: The PORCN inhibitor WNT974 possesses antitumor properties in NET cell lines by inhibiting Wnt and related signaling. In addition, the ß-catenin inhibitor PRI-724 possesses antitumor properties in NET cell lines. Future studies are needed to determine the role of Wnt/ß-catenin signaling in NET as a potential therapeutic target.