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PI3K/Akt and GSK-3β prevents in a differential fashion the malignant phenotype of colorectal cancer cells

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Abstract

Purpose

During colorectal cancer progression, the loss of differentiation and cell–cell adhesion as well as a higher migratory potential are well-defined features; however, the signaling mechanism governing these events is not fully elucidated. The aim of this study was to investigate the role that PI3K and downstream effectors play in controlling colon cancer malignant phenotypes.

Methods

HCT-116 cells, a human model of colon cancer, which are highly metastatic and undifferentiated, were treated with LY294002, a specific inhibitor of PI3K. Cell differentiation and apical junctional complex (AJC) formation were monitored using alkaline phosphatase and electron microscopy analysis. Immunofluorescence and Western blotting were used to accompany the subcellular localization of AJC proteins. PI3K downstream molecules were analyzed by western blotting, and proliferation, wound healing, and colony formation techniques to determine malignant phenotype alterations.

Results

PI3K inhibition increased alkaline phosphatase activity, led to an enterocyte-like growth and formed a functional AJC. LY294002 treatment was able to recruit E-cadherin, β-catenin, claudin-3, and ZO-1 to the cell–cell contact region, and this effect was essential for AJC assembly and association of these proteins to the cytoskeleton. Furthermore, we provided evidence that PI3K inhibition leads to a decrease in p-Akt and p-GSK-3β and increased p-β-catenin levels, which in turn controlled cell proliferation, motility, and colony formation.

Conclusion

Our results demonstrate that PI3K/Akt and GSK-3β prevents in a differential fashion the malignant phenotype of HCT-116 colorectal cancer cells, which could constitute a potential therapeutic target for treatment of this cancer type.

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Acknowledgments

This research was supported by Fundação do Câncer, Ministério da Saúde (MS-Brazil), Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Programa de Cooperação INCA/FIOCRUZ. We thank Dr. Marcelo Pelajo of the Pathology Laboratory, Fiocruz for assistance with confocal microscopy.

Conflict of interest statement

We declare that we have no conflict of interest.

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Authors and Affiliations

  1. Divisão de Biologia Celular, Instituto Nacional de Câncer, Rio de Janeiro, Rua André Cavalcanti 37, 5° andar, 20230-051, Rio de Janeiro, RJ, Brazil

    Wallace Martins de Araújo, Flavia Castello Branco Vidal, Waldemir Fernandes de Souza, Julio César Madureira de Freitas Junior & Jose Andres Morgado-Diaz

  2. Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Ilha do Fundão, 21949-900, Rio de Janeiro, RJ, Brazil

    Wanderley de Souza

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  1. Wallace Martins de Araújo
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  2. Flavia Castello Branco Vidal
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  3. Waldemir Fernandes de Souza
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  6. Jose Andres Morgado-Diaz
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Correspondence to Jose Andres Morgado-Diaz.

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de Araújo, W.M., Vidal, F.C.B., de Souza, W.F. et al. PI3K/Akt and GSK-3β prevents in a differential fashion the malignant phenotype of colorectal cancer cells. J Cancer Res Clin Oncol 136, 1773–1782 (2010). https://doi.org/10.1007/s00432-010-0836-5

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  • DOI: https://doi.org/10.1007/s00432-010-0836-5

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