title: Accelerated biosynthesis of neolacto-series glycosphingolipids in differentiated mouse embryonal carcinoma F9 cells detected by using dodecyl N-acetylglucosaminide as a saccharide primer.
abstract: Using dodecyl N-acetylglucosaminide (GlcNAc-C12) as a saccharide primer, we investigated the biosynthetic changes of neolacto-series glycosphingolipids (GSLs) in mouse embryonal carcinoma F9 cells during differentiation induced by retinoic acid plus dibutyryl cyclic AMP (RA/dbcAMP). In the differentiated cells, the glycosylation of GlcNAc-C12 was greatly enhanced. The sugar compositions of glycosylated primers were assigned as Hex-GlcNAc, [Hex](2)-GlcNAc, [Hex](2)[HexNAc]-GlcNAc, and [NeuAc][Hex]-GlcNAc by liquid chromatography-tandem mass spectrometry. The detection of augmented biosynthesis of endogenous sialylparagloboside indicated that [NeuAc][Hex]-GlcNAc was predicted to be the non-reducing end trisaccharide of sialylparagloboside. The transcription of B3gnt5, B4galt1, Ggta1, Fut4 and St3gal6, encoding glycosyltransferases involved in the neolacto-series glycosphingolipids biosynthesis, was increased, whereas that of Fut9 and St6galI was decreased after RA/dbcAMP treatment. Furthermore, the sialyltransferase activity of ST3GalVI sialylating paragloboside was enhanced with the increase in St3gal6 expression. Since most stage-specific embryonic antigen-1 (SSEA-1) active determinants are carried by glycoproteins in F9 cells, the changes in glycolipid metabolism do not seem to be closely related to loss of cell surface SSEA-1 expression upon F9 differentiation. These results indicate that RA/dbcAMP treatment activates the biosynthesis of neolacto-series GSL and enhances sialylation of paragloboside in F9 cells with down-regulation of Fut9 expression.
authors.affiliation: Department of Pediatric Hematology and Oncology Research, National Research Institute for Child Health and Development, Setagaya-ku, Tokyo, Japan.
authors.forename: Nao
authors.initials: N
authors.lastname: Ogasawara
authors.forename: Yohko U
authors.initials: YU
authors.lastname: Katagiri
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Kiyokawa
authors.forename: Tomonori
authors.initials: T
authors.lastname: Kaneko
authors.forename: Ban
authors.initials: B
authors.lastname: Sato
authors.forename: Hideki
authors.initials: H
authors.lastname: Nakajima
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Miyagawa
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Kushi
authors.forename: Hideharu
authors.initials: H
authors.lastname: Ishida
authors.forename: Makoto
authors.initials: M
authors.lastname: Kiso
authors.forename: Hajime
authors.initials: H
authors.lastname: Okita
authors.forename: Toshinori
authors.initials: T
authors.lastname: Sato
authors.forename: Junichiro
authors.initials: J
authors.lastname: Fujimoto
mesh_terms: D000818:Animals; D015153:Blotting, Western; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002853:Chromatography, Liquid; D002855:Chromatography, Thin Layer; D054278:Embryonal Carcinoma Stem Cells / Q000378:metabolism*; D005434:Flow Cytometry; D006028:Glycosphingolipids / Q000096:biosynthesis*; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053719:Tandem Mass Spectrometry; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D006028:Glycosphingolipids; D014212:Tretinoin; D003994:Bucladesine