The Tissue Architecture of Oral Squamous Cell Carcinoma Visualized by Staining Patterns of Wheat Germ Agglutinin and Structural Proteins Using Confocal Microscopy

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.

Deciphering Cell–Cell Interactions with Integrative Single‐Cell Secretion Profiling

“…[ 36 ] We profiled three types of paired single cells, including SCC25‐CAF (cancer‐associated fibroblasts), OSCC‐CAF, and OSCC‐macrophage, representing tumor–stromal and tumor–immune interactions, respectively. Primary OSCC and CAF cells were isolated from tissues and verified with surface biomarkers immunostaining [ 37 , 38 ] (Figure S4 , Supporting Information). Primary macrophages were derived from a healthy donor.…”

Marine natural product lepadin A as a novel inducer of immunogenic cell death via CD91-dependent pathway

“…2 B). Membrane translocation of CRT on the plasma membrane was visualized by confocal laser scanning microscopy (CLMS) and labelling of the cells with anti-CRT antibody and biotinylated anti-wheat germ agglutinin (WGA) antibody for membrane lectins [ 23 ]. After staining with diamidino-2-phenylindole (DAPI) for nucleic acids, localization of FITC-marked CRT in the cell membranes labeled in red with streptavidin demonstrated the homogeneous distribution of the protein on the cell surface after treatment with lepadin A ( 1 ) and doxorubicin ( 2 ).…”