Human cervical epidermal carcinoma-associated intracellular localization of glycosphingolipid with blood group A type 3 chain

Abstract

A monoclonal antibody, MRG-1, was produced by immunizing a mouse with a human ovarian mucinous cyst adenocarcinoma-derived cell line, RMUG-L. By immunohistochemical staining, the antigen was found to be exclusively localized in the intracellular structures of the cells used as the antigen and of the epithelial cells in normal human cervical glands. However, although the antigen was predominantly detected in the plasma membrane and the intercellular structure of the middle layer of normal human cervical squamous epithelium (92%), it was also contained in the intracellular structure of cervical epidermal carcinoma at a high frequency (80%). The striking difference in the distribution of the MRG-1 antigen between normal and cancerous tissues was found to be a cervical carcinoma-associated phenomenon and a useful tumor marker for immunohistochemical examination. Since the antigen was found to be of a blood group A-related nature by immunohistochemical staining of the tissues and to be a glycosphingolipid, it was purified from human erythrocytes of blood group A, and the structure was concluded to be GalNAc alpha 1-3Gal(2-1 alpha Fuc)beta 1-3GalNAc alpha 1-3Gal(2-1 alpha Fuc)-beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1' Cer, blood group A type 3 chain-containing glycosphingolipid, by NMR, negative ion FABMS and permethylation analysis. In the subcellular localization analysis of the antigen, type 3-A glycosphingolipid antigen was detected in the Golgi body and the microsomes of RMUG-L cells, and the distribution coincided with the finding by immunohistochemical staining. In addition, in cervical epidermal carcinoma, although the blood group A, mainly type 2-A chain, was localized in the plasma membrane and the intercellular structure, the blood group A type 3 chain was selectively found in the perinuclear structure. Also, the blood group A type 3 chain in cervical dysplasia as well as that in normal cervix was predominant in the plasma membrane. Thus, the selective intracellular localization of blood group A type 3 chain was a phenomenon characteristic of cervical epidermal carcinoma and the carcinoma in situ.