Comparison of Total Lipid Composition in Gb3‐Positive and Gb3‐Deficient Burkitt's Lymphoma Cells

Abstract

Thin-layer chromatography was used to compare the glycolipid, phospholipid, and neutral lipid compositions of the globotriaosylceramide (Gb3)‐positive Burkitt's lymphoma Daudi cell line and the Gb3‐deficient Daudi‐derived VT500 cell line. Glycolipid compositions were determined by co‐migration of lipids with glycosphingolipid standards and staining with orcinol. Differences in glycolipid composition of the two cell lines, in addition to Gb3 content, included differences between expression of multiple cerebroside bands and more complex glycolipids. The composition of neutral lipids was similar in the two cell lines. Neutral lipids co‐migrated with triacylglycerol, diacylglycerol, and fatty acid standards. In addition, cholesterol appeared to be the major neutral lipid component in both cell lines. Phospholipid compositions were analyzed by developing chromatograms in two dimensions and staining with molybdenum blue reagent and ninhydrin. As determined by co‐migration with standards and staining properties, phospholipids expressed similarly in both cell lines were identified as lysophosphatidylcholine, lysophosphatidylethanolamine, phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine. Phosphatidic acid and cardiolipin were present in trace amounts. Phospholipids comigrating with phosphatidylinositol and sphingomyelin were strongly expressed in VT500 cells relative to their expression in Daudi cells.

Keywords:

• Thin layer chromatography
• Glycolipids
• Phospholipids
• Neutral lipids
• Burkitt's lymphoma

Acknowledgments

The authors would like to acknowledge grants from NIH/NCMHD/RIMI #MD000215 and NIH/MBRS/SCORE #GM008241 in support of this research and the preparation of this manuscript. We would also like to acknowledge the contributions of Tamu Brown and Veronica Anwuri to earlier research related to this study.