Wnt signaling induces gene expression of factors associated with bone destruction in lung and breast cancer.

Johnson RW, Merkel AR, Page JM, Ruppender NS, Guelcher SA, Sterling JA
Clin Exp Metastasis. 2014 31 (8): 945-59

PMID: 25359619 · PMCID: PMC4258192 · DOI:10.1007/s10585-014-9682-1

Parathyroid hormone-related protein (PTHrP) is an important regulator of bone destruction in bone metastatic tumors. Transforming growth factor-beta (TGF-β) stimulates PTHrP production in part through the transcription factor Gli2, which is regulated independent of the Hedgehog signaling pathway in osteolytic cancer cells. However, inhibition of TGF-β in vivo does not fully inhibit tumor growth in bone or tumor-induced bone destruction, suggesting other pathways are involved. While Wnt signaling regulates Gli2 in development, the role of Wnt signaling in bone metastasis is unknown. Therefore, we investigated whether Wnt signaling regulates Gli2 expression in tumor cells that induce bone destruction. We report here that Wnt activation by β-catenin/T cell factor 4 (TCF4) over-expression or lithium chloride (LiCl) treatment increased Gli2 and PTHrP expression in osteolytic cancer cells. This was mediated through the TCF and Smad binding sites within the Gli2 promoter as determined by promoter mutation studies, suggesting cross-talk between TGF-β and Wnt signaling. Culture of tumor cells on substrates with bone-like rigidity increased Gli2 and PTHrP production, enhanced autocrine Wnt activity and led to an increase in the TCF/Wnt signaling reporter (TOPFlash), enriched β-catenin nuclear accumulation, and elevated Wnt-related genes by PCR-array. Stromal cells serve as an additional paracrine source of Wnt ligands and enhanced Gli2 and PTHrP mRNA levels in MDA-MB-231 and RWGT2 cells in vitro and promoted tumor-induced bone destruction in vivo in a β-catenin/Wnt3a-dependent mechanism. These data indicate that a combination of matrix rigidity and stromal-secreted factors stimulate Gli2 and PTHrP through Wnt signaling in osteolytic breast cancer cells, and there is significant cross-talk between the Wnt and TGF-β signaling pathways. This suggests that the Wnt signaling pathway may be a potential therapeutic target for inhibiting tumor cell response to the bone microenvironment and at the very least should be considered in clinical regimens targeting TGF-β signaling.

MeSH Terms (26)

Animals beta Catenin Blotting, Western Bone Neoplasms Breast Neoplasms Female Gene Expression Regulation Humans Kruppel-Like Transcription Factors Lung Neoplasms Mice Mice, Nude Nuclear Proteins Parathyroid Hormone-Related Protein Promoter Regions, Genetic Real-Time Polymerase Chain Reaction Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger RNA, Small Interfering Signal Transduction Stromal Cells Transforming Growth Factor beta Tumor Cells, Cultured Wnt3A Protein Xenograft Model Antitumor Assays Zinc Finger Protein Gli2

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