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Issue 6, 2012
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The lipid composition of isolated cytoplasmic lipid droplets from a human cancer cell line, BE(2)M17

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Abstract

1H nuclear magnetic resonance spectroscopy (NMR) resonances from lipids in tumours are associated with tumour grade and treatment response. The origin of these NMR signals is mainly considered to be cytoplasmic lipid droplets (LDs). Techniques exist for isolating LDs but little is known about their composition and its relationship to NMR signals. In this work, density-gradient ultracentrifugation was performed on homogenised human cancer cells to isolate LDs. 1H NMR was performed on whole cells, isolated LDs and their extracts. Heteronuclear single quantum coherence spectroscopy (HSQC) and liquid chromatography mass spectroscopy (LC-MS) were performed on lipid extracts of LDs. Staining and microscopy were used to characterize isolated LDs. An excellent agreement in chemical shift and relative signal intensity was observed between lipid resonances in cells and isolated LD spectra supporting that NMR-visible lipids originate primarily from LDs. Isolated LDs showed high concentrations of unsaturated lipids, a oleic-to-linoleic acid ratio greater than two and a cholesteryl ester (ChE)-to-cholesterol (Ch) ratio close to unity. These ratios were several-fold greater than respective ratios in whole cells, demonstrating isolation is important to characterize LD composition. LDs contain a specific group of lipid species that are likely to contribute to the 1H NMR spectrum of cells.

Graphical abstract: The lipid composition of isolated cytoplasmic lipid droplets from a human cancer cell line, BE(2)M17

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Publication details

The article was received on 24 Nov 2011, accepted on 16 Mar 2012, published on 20 Apr 2012 and first published online on 20 Apr 2012


Article type: Paper
DOI: 10.1039/C2MB05485J
Citation: Mol. BioSyst., 2012,8, 1694-1700
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    The lipid composition of isolated cytoplasmic lipid droplets from a human cancer cell line, BE(2)M17

    X. Pan, M. Wilson, C. McConville, M. Brundler, T. N. Arvanitis, J. P. Shockcor, J. L. Griffin, R. A. Kauppinen and A. C. Peet, Mol. BioSyst., 2012, 8, 1694
    DOI: 10.1039/C2MB05485J

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