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Originally published as JHC exPRESS on August 9, 2006.
doi:10.1369/jhc.6A6958.2006
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Journal of Histochemistry and Cytochemistry
Volume 54 (11): 1263-1275, 2006
Copyright ©The Histochemical Society, Inc.

Snail-dependent and -independent Epithelial–Mesenchymal Transition in Oral Squamous Carcinoma Cells

Minna Takkunen, Reidar Grenman, Mika Hukkanen, Matti Korhonen, Antonio García de Herreros and Ismo Virtanen

Institute of Biomedicine/Anatomy, University of Helsinki, Helsinki, Finland (MT,MH,IV); Department of Otorhinolaryngology, Head and Neck Surgery, Turku University Central Hospital, Turku, Finland (RG); Hospital for Children and Adolescents, Helsinki University Central Hospital, Helsinki, Finland (MK); and Unitat de Biologia Cel.lular i Molecular, Institut Municipal d'Investigació Mèdica, Universitat Pompeu Fabra, Barcelona, Spain (AGH)

Correspondence to: Minna Takkunen, MD, Institute of Biomedicine/Anatomy, P.O. Box 63 (Haartmaninkatu 8), FI-00014 University of Helsinki, Helsinki, Finland. E-mail: minna.k.takkunen{at}helsinki.fi

Disappearance of E-cadherin is a milestone for epithelial–mesenchymal transition (EMT), found both in carcinomas and in some fibrotic diseases. We have studied the mechanisms of EMT in oral squamous cell carcinoma (SCC) cells isolated from primary tumor (43A) and its recurrent tumor (43B). Whereas the cells from primary carcinoma displayed a typical phenotype of squamous epithelial cells including E-cadherin and laminin-332 (laminin-5), cells from recurrent tumor expressed characteristics of dedifferentiated, EMT-experienced tumors. 43B cells expressed E-cadherin repressors ZEB-1/{delta}EF1 and especially ZEB-2/SIP1, which therefore appear as candidates for endogenous EMT in these cells. Differences between endogenous and exogenous EMT were assessed by transfecting 43A cells with SNAIL cDNA. SNAIL-transfected cells showed complete EMT phenotype with fibroblastoid appearance, vimentin filaments, E-cadherin/N-cadherin switch, lack of hemidesmosomes and, as a new feature of EMT, lack of laminin-332 synthesis. Upregulation of ZEB-1 and ZEB-2 was evident in these cells, suggesting that SNAIL can regulate these E-cadherin repressors. New monoclonal antibodies against SNAIL showed nuclear immunoreactivity not only in the SNAIL-transfected cells but also in carcinoma cells lacking production of Lm-332 and showing signs of EMT. These results suggest that changes in the epithelial cell differentiation program and EMT in SCC cells can result from the interplay among several E-cadherin repressors; however, SNAIL alone is able to accomplish a complete EMT. (J Histochem Cytochem 54:1263–1275, 2006)

Key Words: epithelial–mesenchymal transition • E-cadherin • ZEB-1 • ZEB-2 • SNAIL • laminin-332


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