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Abstract
Objectives: Tissue architecture and cell morphology suffer profound alterations during oral cancer and are important markers for its progression and outcome. For precise visualization of tissue architecture in oral cancer, we used confocal microscopy to examine the staining pattern of wheat germ agglutinin, a lectin that binds membrane glycoproteins, and the staining patterns of structural proteins. Materials and Methods: Paraffin sections of oral squamous cell carcinoma were stained with fluorescently labeled wheat germ agglutinin and with antibodies against structural proteins, which were revealed by immunohistochemistry with tyramide signal amplification. Results: Membrane localization of wheat germ agglutinin was markedly decreased in the basal layers and in regions of tumor invasion, accompanied by cytoplasmic redistribution of E-cadherin, β-actin and syndecan-1. Wheat germ agglutinin staining clearly identified tumor clusters within the surrounding stroma, and tumor cells with elongated morphology. Conclusions: Our results suggest that the wheat germ agglutinin staining pattern is indicative of the degree of cell cohesion in oral squamous cell carcinoma, which decreases in basal layers and invasive tumor clusters with more migratory morphologies. Wheat germ agglutinin staining in combination with confocal microscopy could constitute, therefore, a valuable tool for the study of tissue architecture in oral cancer.
Highlights
Tumor growth and its progressive taking over of neighboring tissues impose drastic alterations in tissue architecture, which have prognostic value in many types of cancers [1,2], including oral squamous cell carcinoma (OSCC)
Wheat germ agglutinin (WGA) is a plant lectin with high affinity for N-acetylglucosamine and sialic acid [7], which can bind to multiple membrane glycoproteins containing these monosaccharides in their varying carbohydrate moieties, and in particular it can bind to cell adhesion molecules [8,9,10]
Altered tissue architecture, especially at the level of cell–cell adhesion, produces a clearly altered pattern of WGA staining [11]. Another important consideration for studies of tumor architecture with high resolution microscopy is the frequently mentioned difficulty of performing immunofluorescence staining in formalin-fixed, paraffin embedded (FFPE) thin sections [13], which is relevant given the prevalence of archival FFPE tumor samples, but this can be circumvented, for example, by the use of signal amplification techniques such as tyramide signal amplification (TSA), which generates covalently bound fluorophores in very close proximity to the detected epitope [14]
Summary
Tumor growth and its progressive taking over of neighboring tissues impose drastic alterations in tissue architecture, which have prognostic value in many types of cancers [1,2], including oral squamous cell carcinoma (OSCC). Altered tissue architecture, especially at the level of cell–cell adhesion, produces a clearly altered pattern of WGA staining [11] Another important consideration for studies of tumor architecture with high resolution microscopy is the frequently mentioned difficulty of performing immunofluorescence staining in formalin-fixed, paraffin embedded (FFPE) thin sections [13], which is relevant given the prevalence of archival FFPE tumor samples, but this can be circumvented, for example, by the use of signal amplification techniques such as tyramide signal amplification (TSA), which generates covalently bound fluorophores in very close proximity to the detected epitope [14]
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