Glycobiology Advance Access originally published online on June 18, 2009
Glycobiology 2009 19(9):1018-1033; doi:10.1093/glycob/cwp083
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Unusual accumulation of sulfated glycosphingolipids in colon cancer cells
2 Department of Immunology, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-2 Nakamichi, Higashinari-ku, Osaka 537-8511
3 Department of Disease Glycomics, The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047
4 Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, 1-3-3 Nakamichi, Higashinari-ku, Osaka 537-8511
5 Department of Biochemistry
6 Kochi System Glycobiology Center, Kochi University Medical School, Nankoku, Kochi 783-8505
7 CREST, Japan Science and Technology Agency, Kawaguchi, Japan
1 To whom correspondence should be addressed: Tel: +81-6-6972-1181; Fax: +81-6-6972-7749; e-mail: miyamoto-ya{at}mc.pref.osaka.jp
Received on April 30, 2009; revised on June 5, 2009; accepted on June 8, 2009
The structures of glycosphingolipids from highly purified colorectal cancer cells and normal colorectal epithelial cells of 16 patients have been analyzed in fine detail (Misonou Y, Shida K, Korekane H, Seki Y, Noura S, Ohue M, Miyamoto Y. 2009. Comprehensive Clinico-Glycomic Study of 16 Colorectal Cancer Specimens: Elucidation of aberrant glycosylation and ts mechanistic causes in colorectal cancer cells. J Proteome Res. 8:2990–3005). Further structural analyses demonstrated that colon cancer cells from two patients accumulated unusual glycosphingolipids which were not observed in either colorectal cancer cells or normal colorectal epithelial cells from the other patients. Mass spectrometry analyses revealed that the unusual structures include sulfated oligosaccharides. The structures of the glycosphingolipids of the cancer cells from these two cases were analyzed by methods which include enzymatic release of carbohydrate moieties, fluorescent labeling with aminopyridine and identification using two-dimensional mapping, enzymatic digestion and mass spectrometry together with methanolysis, and the use of newly synthesized sulfo-fucosylated oligosaccharides as standards. The colon cancer cells from one of the patients demonstrate a variety of oligosaccharides as major components which are sulfated at the C6 position of subterminal GlcNAc and at C3 positions of terminal galactose with or without sialylation or fucosylation. These include 6-sulfo Lex, 6'-sialyl 6-sulfo lactosamine, and 3'-sialyl 6-sulfo Lex, in addition to sialylated or fucosylated derivatives of type-1 and type-2 hybrid oligosaccharides. The colon cancer cells from the other patient have two kinds of sulfated oligosaccharides, a 6-sulfo Lex structure and a 3'-sulfo Lex structure, as minor components. Taking into consideration the clinical features of the two patients, the biological significance of sulfated glycosphingolipids on cancer cells is discussed.
Key words: colon cancer / glycosphingolipid / sulfate / two-dimensional mapping