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Wnt-5a has tumor suppressor activity in thyroid carcinoma
N Kremenevskaja1, R von Wasielewski, A S Rao
1Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Carl-Neuberg Str. 1, D-30625 Hannover, Germany.
Abstract:
Stabilization of beta-catenin by inhibition of its phosphorylation is characteristic of an activation of the canonical Wnt/beta-catenin signaling pathway and is associated with various human carcinomas. It contrasts to an as yet incompletely characterized action of an alternative noncanonical Wnt signaling pathway on neoplastic transformation. The aim of the present study was to test the effects of a member of the noncanonical Wnt signaling pathway, Wnt-5a, in primary thyroid carcinomas and in thyroid carcinoma cell lines. Compared to normal tissue Wnt-5a mRNA expression was clearly increased in thyroid carcinomas. Immunohistochemically, a bell-shaped response was observed with low to undetectable levels in normal tissue and in anaplastic tumors whereas differentiated thyroid carcinomas showed strong positive immunostaining for Wnt-5a. Transfection of Wnt-5a in a thyroid tumor cell line FTC-133 was able to reduce proliferation, migration, invasiveness and clonogenicity in these cells. These effects of Wnt-5a are associated with membranous beta-catenin translocation and c-myc oncogene suppression and are mediated through an increase in intracellular Ca(2+) release, which via CaMKII pathways promotes beta-catenin phosphorylation. Specific inhibition of beta-catenin phosphorylation by W-7, a calmodulin inhibitor, or by KN-93, a CaMKII inhibitor, supports these findings whereas PKC inhibitors were without effect. This interaction occurs downstream of GSK-3 beta as no Wnt-5a effect was seen on the Ser(9) phosphorylation of GSK-3 beta. Our data are compatible with the hypothesis that Wnt-5a serves as an antagonist to the canonical Wnt-signaling pathway with tumor suppressor activity in differentiated thyroid carcinomas.
Insights
Wnt-5a, a noncanonical Wnt pathway molecule, acts as a tumor suppressor in differentiated thyroid carcinomas. It inhibits proliferation and invasiveness by promoting beta-catenin phosphorylation, opposing canonical Wnt/beta-catenin signaling.
Area of Science:
- Oncology
- Molecular Biology
- Cell Signaling
Background:
- Canonical Wnt/beta-catenin signaling pathway activation, marked by beta-catenin stabilization, is linked to human carcinomas.
- Noncanonical Wnt signaling pathways also influence neoplastic transformation, but their roles are less understood.
- Wnt-5a is a key member of the noncanonical Wnt signaling pathway.
Purpose of the Study:
- To investigate the effects of Wnt-5a in primary thyroid carcinomas and cell lines.
- To determine the role of Wnt-5a in thyroid tumor development and progression.
Main Methods:
- Quantitative analysis of Wnt-5a mRNA expression in normal thyroid tissue and thyroid carcinomas.
- Immunohistochemical analysis of Wnt-5a protein expression in thyroid tumors.
- Wnt-5a gene transfection into FTC-133 thyroid tumor cells to assess proliferation, migration, and invasiveness.
- Analysis of beta-catenin and GSK-3 beta phosphorylation, intracellular calcium release, and downstream signaling pathways (CaMKII, PKC).
Main Results:
- Wnt-5a mRNA expression was significantly increased in thyroid carcinomas compared to normal tissue.
- Immunohistochemistry revealed a bell-shaped expression pattern: low in normal tissue and anaplastic tumors, high in differentiated thyroid carcinomas.
- Wnt-5a transfection reduced proliferation, migration, invasiveness, and clonogenicity of FTC-133 cells.
- Wnt-5a induced membranous beta-catenin translocation, c-myc suppression, and increased intracellular Ca(2+) release, promoting beta-catenin phosphorylation via CaMKII.
- Inhibition of beta-catenin phosphorylation by W-7 or KN-93 confirmed the CaMKII-dependent mechanism, while PKC inhibitors had no effect.
- Wnt-5a did not affect GSK-3 beta phosphorylation at Ser(9), indicating downstream action.
Conclusions:
- Wnt-5a exhibits tumor suppressor activity in differentiated thyroid carcinomas.
- Wnt-5a antagonizes the canonical Wnt/beta-catenin signaling pathway through promoting beta-catenin phosphorylation via a Ca(2+)/CaMKII-dependent mechanism.
- These findings highlight Wnt-5a as a potential therapeutic target in thyroid cancer.
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