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Claudin-1 overexpression in melanoma is regulated by PKC and contributes to melanoma cell motility

Oncogene volume 26, pages 3846–3856 (2007)Cite this article

Abstract

Serial analysis of gene expression followed by pathway analysis implicated the tight junction protein claudin-1 (CLDN1) in melanoma progression. Tight junction proteins regulate the paracellular transport of molecules, but staining of a tissue microarray revealed that claudin-1 was overexpressed in melanoma, and aberrantly expressed in the cytoplasm of malignant cells, suggesting a role other than transport. Indeed, melanoma cells in culture demonstrate no tight junction function. It has been shown that protein kinase C (PKC) can affect expression of claudin-1 in rat choroid plexus cells, and we observed a correlation between levels of activated PKC and claudin expression in our melanoma cells. To determine if PKC could affect the expression of CLDN1 in human melanoma, cells lacking endogenous claudin-1 were treated with 200 nM phorbol myristic acid (PMA). PKC activation by PMA caused an increase in CLDN1 transcription in 30 min, and an increase in claudin-1 protein by 12 h. Inhibition of PKC signaling in cells with high claudin-1 expression resulted in decreased claudin-1 expression. CLDN1 appears to contribute to melanoma cell invasion, as transient transfection of melanoma cells with CLDN1 increased metalloproteinase 2 (MMP-2) secretion and activation, and subsequently, motility of melanoma cells as demonstrated by wound-healing assays. Conversely, knockdown of CLDN1 by siRNA resulted in the inhibition of motility, as well as decreases in MMP-2 secretion and activation. These data implicate claudin-1 in melanoma progression.

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Acknowledgements

We thank Drs Michel Bernier, Vishwa Deep-Dixit, Teresa D'Souza and Rachana Agarwal for helpful discussion, reagents and technical advice. We thank Dr Meenhard Herlyn for his generous gift of cell lines, and Ms Ana Lustig for help with the pathway analysis software. We thank Drs Jeff Trent, Michael Bittner, Galen Hostetter and Urs Wagner for the melanoma tissue microarray. We also thank Mr Jason Godlove for help with statistical analysis. This work was supported by funding from the Intramural Research Program of the National Institute on Aging, National Institutes of Health, Baltimore, MD, USA.

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Author notes

  1. M J Rhode

    Present address: Nucleonics, Delaware Valley College, DE, USA

  2. H F Brown

    Present address: Purdue University, West Lafayette, IN, USA

Authors and Affiliations

  1. Laboratory of Immunology, Gerontology Research Center, National Institute on Aging, Baltimore, MD, USA

    P D Leotlela, M S Wade, M J Rhode, H F Brown, D T Rosenthal, S K Dissanayake, D D Taub & A T Weeraratna

  2. VA Healthcare System, Boston, MA, USA

    P H Duray

  3. MARC scholar, University of Maryland Eastern Shore, Princess Anne, MD, USA

    H F Brown

  4. Department of Biology and Marine Biology, University of North Carolina, Wilmington, NC, USA

    D T Rosenthal

  5. Research Resources Branch, Gerontology Research Center, National Institute on Aging, Baltimore, MD, USA

    R Earley & F E Indig

  6. Department of Pathology, Loyola University Medical Center, Maywood, IL, USA

    B J Nickoloff

  7. Medical Biotechnology Department, VTT Technical Research Centre of Finland, University of Turku, Turku, Finland

    O P Kallioniemi

  8. Cancer Genetics Branch, National Human Genome Research Institute, Bethesda, MD, USA

    P Meltzer

  9. Laboratory of Cellular and Molecular Biology, Gerontology Research Center, National Institute on Aging, Baltimore, MD, USA

    P J Morin

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Correspondence to A T Weeraratna.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Leotlela, P., Wade, M., Duray, P. et al. Claudin-1 overexpression in melanoma is regulated by PKC and contributes to melanoma cell motility. Oncogene 26, 3846–3856 (2007). https://doi.org/10.1038/sj.onc.1210155

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