Expression and localisation of MUC1 modified with sialylated core-2 O-glycans in mucoepidermoid carcinoma
Takanori Sugiura, Kazuhiko Hashimoto, Kazutaka Kikuta, Ukei Anazawa, Takeshi Nomura, Akihiko Kameyama
Abstract
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Takanori Sugiura, Kazuhiko Hashimoto, Kazutaka Kikuta, Ukei Anazawa, Takeshi Nomura, Akihiko Kameyama
Abstract
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Abstract Mucoepidermoid carcinoma (MEC) is the most frequent of the rare salivary gland malignancies. We previously reported high expression of Mucin 1 (MUC1) modified with sialylated core-2 O-glycans in MEC by using tissue homogenates. In this study, we characterised glycan structures of MEC and identified the localisation of cells expressing these distinctive glycans on MUC1. Mucins were extracted from the frozen tissues of three patients with MEC, and normal salivary glands (NSGs) were extracted from seven patients, separated by supported molecular matrix electrophoresis (SMME) and the membranes stained with various lectins. In addition, formalin-fixed, paraffin-embedded sections from three patients with MEC were subjected to immunohistochemistry (IHC) with various monoclonal antibodies and analysed for C2GnT-1 expression by in situ hybridisation (ISH). Lectin blotting of the SMME membranes revealed that glycans on MUC1 from MEC samples contained α2,3-linked sialic acid. In IHC, MUC1 was diffusely detected at MEC-affected regions but was specifically detected at apical membranes in NSGs. ISH showed that C2GnT-1 was expressed at the MUC1-positive in MEC-affected regions but not in the NSG. Conclusion: MEC cells produced MUC1 modified with α2,3-linked sialic acid-containing core 2 O-glycans. MUC1 containing these glycans deserves further study as a new potential diagnostic marker of MEC. (200 words)
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Abstract Mucoepidermoid carcinoma (MEC) is the most frequent of the rare salivary gland malignancies. We previously reported high expression of Mucin 1 (MUC1) modified with sialylated core-2 O-glycans in MEC by using tissue homogenates. In this study, we characterised glycan structures of MEC and identified the localisation of cells expressing these distinctive glycans on MUC1. Mucins were extracted from the frozen tissues of three patients with MEC, and normal salivary glands (NSGs) were extracted from seven patients, separated by supported molecular matrix electrophoresis (SMME) and the membranes stained with various lectins. In addition, formalin-fixed, paraffin-embedded sections from three patients with MEC were subjected to immunohistochemistry (IHC) with various monoclonal antibodies and analysed for C2GnT-1 expression by in situ hybridisation (ISH). Lectin blotting of the SMME membranes revealed that glycans on MUC1 from MEC samples contained α2,3-linked sialic acid. In IHC, MUC1 was diffusely detected at MEC-affected regions but was specifically detected at apical membranes in NSGs. ISH showed that C2GnT-1 was expressed at the MUC1-positive in MEC-affected regions but not in the NSG. Conclusion: MEC cells produced MUC1 modified with α2,3-linked sialic acid-containing core 2 O-glycans. MUC1 containing these glycans deserves further study as a new potential diagnostic marker of MEC. (200 words)
Key concepts: MUC1, Mucin, Sialic acid, Immunohistochemistry, Glycan, Mucoepidermoid carcinoma, Molecular biology, Lectin