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PKG inhibits TCF signaling in colon cancer cells by blocking β-catenin expression and activating FOXO4

Oncogene volume 29, pages 3423–3434 (2010)Cite this article

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Abstract

Activation of cGMP-dependent protein kinase (PKG) has anti-tumor effects in colon cancer cells but the mechanisms are not fully understood. This study has examined the regulation of β-catenin/TCF signaling, as this pathway has been highlighted as central to the anti-tumor effects of PKG. We show that PKG activation in SW620 cells results in reduced β-catenin expression and a dramatic inhibition of TCF-dependent transcription. PKG did not affect protein stability, nor did it increase phosphorylation of the amino-terminal Ser33/37/Thr41 residues that are known to target β-catenin for degradation. However, we found that PKG potently inhibited transcription from a luciferase reporter driven by the human CTNNB1 promoter, and this corresponded to reduced β-catenin mRNA levels. Although PKG was able to inhibit transcription from both the CTNNB1 and TCF reporters, the effect on protein levels was less consistent. Ectopic PKG had a marginal effect on β-catenin protein levels in SW480 and HCT116 but was able to inhibit TCF-reporter activity by over 80%. Investigation of alternative mechanisms revealed that cJun-N-terminal kinase (JNK) activation was required for the PKG-dependent regulation of TCF activity. PKG activation caused β-catenin to bind to FOXO4 in colon cancer cells, and this required JNK. Activation of PKG was also found to increase the nuclear content of FOXO4 and increase the expression of the FOXO target genes MnSOD and catalase. FOXO4 activation was required for the inhibition of TCF activity as FOXO4-specific short-interfering RNA completely blocked the inhibitory effect of PKG. These data illustrate a dual-inhibitory effect of PKG on TCF activity in colon cancer cells that involves reduced expression of β-catenin at the transcriptional level, and also β-catenin sequestration by FOXO4 activation.

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Acknowledgements

Supported by grants from the American Cancer Society (RSG-07-174-01-CSM to DDB and RSG-09-209-01-TBG to KL) and the National Institutes of Health (CA133085 to KL).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta, GA, USA

    I-K Kwon, R Wang, M Thangaraju, H Shuang, K Liu, V Ganapathy & D D Browning

  2. Cancer Chemoprotection Program, Linus Pauling Institute, Oregon State University, Corvallis, OR, USA

    R Dashwood

  3. Department of Medicine, Section of Pulmonary and Critical Care Medicine, University of Chicago, Chicago, IL, USA

    N Dulin

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Kwon, IK., Wang, R., Thangaraju, M. et al. PKG inhibits TCF signaling in colon cancer cells by blocking β-catenin expression and activating FOXO4. Oncogene 29, 3423–3434 (2010). https://doi.org/10.1038/onc.2010.91

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