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BMC Cancer
Published in: BMC Cancer 1/2017

Open Access 01-12-2017 | Research article

A novel FOXO1-mediated dedifferentiation blocking role for DKK3 in adrenocortical carcinogenesis

Authors: Joyce Y. Cheng, Taylor C. Brown, Timothy D. Murtha, Adam Stenman, C. Christofer Juhlin, Catharina Larsson, James M. Healy, Manju L. Prasad, Wolfram T. Knoefel, Andreas Krieg, Ute I. Scholl, Reju Korah, Tobias Carling

Published in: BMC Cancer | Issue 1/2017

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Abstract

Background

Dysregulated WNT signaling dominates adrenocortical malignancies. This study investigates whether silencing of the WNT negative regulator DKK3 (Dickkopf-related protein 3), an implicated adrenocortical differentiation marker and an established tumor suppressor in multiple cancers, allows dedifferentiation of the adrenal cortex.

Methods

We analyzed the expression and regulation of DKK3 in human adrenocortical carcinoma (ACC) by qRT-PCR, immunofluorescence, promoter methylation assay, and copy number analysis. We also conducted functional studies on ACC cell lines, NCI-H295R and SW-13, using siRNAs and enforced DKK3 expression to test DKK3’s role in blocking dedifferentiation of adrenal cortex.

Results

While robust expression was observed in normal adrenal cortex, DKK3 was down-regulated in the majority (>75%) of adrenocortical carcinomas (ACC) tested. Both genetic (gene copy loss) and epigenetic (promoter methylation) events were found to play significant roles in DKK3 down-regulation in ACCs. While NCI-H295R cells harboring β-catenin activating mutations failed to respond to DKK3 silencing, SW-13 cells showed increased motility and reduced clonal growth. Conversely, exogenously added DKK3 also increased motility of SW-13 cells without influencing their growth. Enforced over-expression of DKK3 in SW-13 cells resulted in slower cell growth by an extension of G1 phase, promoted survival of microcolonies, and resulted in significant impairment of migratory and invasive behaviors, largely attributable to modified cell adhesions and adhesion kinetics. DKK3-over-expressing cells also showed increased expression of Forkhead Box Protein O1 (FOXO1) transcription factor, RNAi silencing of which partially restored the migratory proficiency of cells without interfering with their viability.

Conclusions

DKK3 suppression observed in ACCs and the effects of manipulation of DKK3 expression in ACC cell lines suggest a FOXO1-mediated differentiation-promoting role for DKK3 in the adrenal cortex, silencing of which may allow adrenocortical dedifferentiation and malignancy.
Appendix
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Metadata
Title
A novel FOXO1-mediated dedifferentiation blocking role for DKK3 in adrenocortical carcinogenesis
Authors
Joyce Y. Cheng
Taylor C. Brown
Timothy D. Murtha
Adam Stenman
C. Christofer Juhlin
Catharina Larsson
James M. Healy
Manju L. Prasad
Wolfram T. Knoefel
Andreas Krieg
Ute I. Scholl
Reju Korah
Tobias Carling
Publication date
01-12-2017
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2017
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-017-3152-5

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