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Individual profiles of free ceramide species and the constituent ceramide species of sphingomyelin and neutral glycosphingolipid and their alteration according to the sequential changes of environmental oxygen content in human colorectal cancer Caco-2 cells

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Abstract

We previously performed a systematic analysis of free ceramide (Cers) species, the constituent ceramide species of sphingomyelins and neutral glycosphingolipids (NGSLs) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry with high-energy collision-induced dissociation. As a result, distinct species differences were found among Cers, sphingomyelins and NGSLs in the kidneys. Using this method, we investigated various sphingolipid species from human colon cancer Caco-2 cells as well as the influence of environmental oxygen on these species in detail. Unexpectedly, even in normoxia, all Cers species were composed of dihydrosphingosine (d18:0) and non-hydroxy fatty acid (NFA), and 34 % of sphingomyelins were composed of dihydrosphingomyelins with NFA. In contrast, major constituent ceramide species of NGSLs were composed of the usual long-chain base of sphingosine (d18:1) and hydroxy fatty acid (HFA). When the cells were cultured under hypoxic condition for 3 days, all the Cers and nearly 80 % of the sphingomyelins were dihydrosphingolipids composed of d18:0-NFAs, but a significant proportion of d18:1-HFAs still remained in the NGSLs. When the cells were transferred from conditions of hypoxia to normoxia again (reoxygenation), Cer species composed of d18:1-NFAs, which were not found in Cers under the original normoxic conditions, appeared. Such Cers were probably synthesized as precursors for the constituent ceramides of sphingomyelins and NGSLs.

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Authors and Affiliations

  1. Department of Medical Technology, Nagoya University Graduate School of Health Sciences, Daiko-Minami 1-1-20, Higashi-ku, Nagoya, 461-8673, Japan

    Kouji Tanaka, Keiko Tamiya-Koizumi & Takashi Murate

  2. Shimadzu Corporation, Nishinokyo-Kuwabaracho 1, Nakagyo-ku, Kyoto, 604-8511, Japan

    Masaki Yamada

  3. Research Complex for the Medicine Frontiers, Aichi Medical University, Yazako, Nagakute, Aichi, 480-1195, Japan

    Reiji Kannagi

  4. Department of Microbiology and Molecular Cell Biology, Nihon Pharmaceutical University, Komuro 10281, Inamachi, Saitama, 362-0806, Japan

    Mamoru Kyogashima

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  1. Kouji Tanaka
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  2. Keiko Tamiya-Koizumi
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Correspondence to Mamoru Kyogashima.

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Supplemental Fig. 1

Fragmentation scheme and MS/MS profile of the [M + H]+ ion at m/z 540.2 derived from Cer of d18:0-C16:0 and its spectrum (a). MS/MS profile of the [M + H] + ion at m/z 789.9 derived from sphingomyelin d18:0-C22:0. Ion at m/z 184.5 derived from phosphocholine (b). (PDF 187 kb)

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Tanaka, K., Tamiya-Koizumi, K., Yamada, M. et al. Individual profiles of free ceramide species and the constituent ceramide species of sphingomyelin and neutral glycosphingolipid and their alteration according to the sequential changes of environmental oxygen content in human colorectal cancer Caco-2 cells. Glycoconj J 31, 209–219 (2014). https://doi.org/10.1007/s10719-013-9511-9

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  • DOI: https://doi.org/10.1007/s10719-013-9511-9

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