Glycosphingolipids of human KB cells grown in monolayer, suspension, and synchronized cultures

Studies have been carried out on the glycosphingolipids of human KB cells grown in monolayer and suspension culture, and by synchronization of the latter with a double thymidine (2mM) block. Glycosphingolipids were identified tentatively by thin layer chromatography, gas-liquid chromatography, and c...

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Bibliographic Details
Published in:The Journal of biological chemistry 1975-01, Vol.250 (1), p.61-66
Main Authors: Chatterjee, S, Sweeley, C C, Velicer, L F
Format: Article
Language:English
Subjects:
Carbon Radioisotopes
Carcinoma
Cell Division
Cell Line
Chromatography, Gas
Chromatography, Thin Layer
Culture Techniques - methods
Galactosamine - analysis
Galactose - analysis
Galactose - metabolism
Gangliosides - biosynthesis
Glucose - analysis
Glycolipids - biosynthesis
Humans
Mass Spectrometry
Mouth Neoplasms
Sialic Acids - analysis
Sphingolipids - biosynthesis
Thymidine - metabolism
Time Factors
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Summary:Studies have been carried out on the glycosphingolipids of human KB cells grown in monolayer and suspension culture, and by synchronization of the latter with a double thymidine (2mM) block. Glycosphingolipids were identified tentatively by thin layer chromatography, gas-liquid chromatography, and combined gas-liquid chromatography and mass spectrometry. The predominant gangliosides in the these cells were AcNeu-Gal-Glc-Cer and AcNeu-Gal-GalNAc-Gal-(AcNeu). Glc-Cer. Theprincipal neutral glycosphingolipids were Glc-Cer, Gal-Glc-Cer, Gal-Gal-Glc-Cer, and GalNAc-Gal-Gal-Glc-Cer. Incubation of KB cells (grown in monolayer and subsequently in suspension culture) for 48 hours with D-[1-14Clgalactose resulted in appreciable incorporation of radioactivity into all of the principal glycosphingolipids of these cells. These experiments confirmed that KB cells are capable of synthesizing their constituent glycosphingolipids. KB cells grown in suspension culture showed A 2- to 3-fold increase inthe concentration of Glc-Cer, Gal-Glc-Cer, GalNAc-Gal-Gal-Glc-Cer, and AcNeu-Gal1NAc-Gal-Gal-Glc-Cer, and AcNeu-Gal-Ga1NAcGal-(AcNeu)-Glc-Cer. Thus, the occurrence of tissue culture-dependent changes in the level of glycosphingolipids is demonstrated. Perhaps messages governing the synthesis of glycosphingolipids are translated earlier in thecell cycle under certain conditions of growth and are affected by cell-cell contact and cell adhesion.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)41979-0