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Effect of eicosapentaenoic acid on the proliferation and incidence of apoptosis in the colorectal cell line HT29

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Lipids

Abstract

Fish oil has been shown to reduce the induction of colorectal cancer in animal models by a mechanism which may involve suppression of mitosis, increased apoptosis, or both. We used the human colonic adenocarcinoma cell line HT29 to explore the effects of the long-chain n-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA) on cell proliferation and death in vitro. Cells were cultured in media containing EPA at 5, 10, and 15 μg/mL. Cell number and thymidine incorporation were used to quantify proliferation, and cell cycle effects were studied using flow cytometry. Gel electrophoresis, annexin-V binding, and morphological criteria were used to characterize apoptosis. Adherent cells and freely floating detached cells were treated as two distinct populations. In the presence of EPA at 10 and 15 μg/mL there was a marked reduction in the growth rate of adherent HT29 colonies, owing to an increased detachment of adherent cells. After treatment with 10 or 15 μg/mL EPA the proportion of adherent cells in S-phase increased, indicating either a block in late S-phase or early G2. Floating cells showed evidence of extensive DNA cleavage, but the proportion of floating cells with sub G0 DNA content declined on treatment with 10 or 15 μg/mL EPA even though the number of floating cells increased. We conclude that EPA does not inhibit mitosis of adherent cells, but increases the rate at which they become detached from the substrate, probably at an early stage in the initiation of apoptosis. This mechanism may be analogous to “anoikis”, or induction of apoptosis in response to loss of cell contact, and may contribute to the anticarcinogenic effects of fish oil in vivo.

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Abbreviations

DMEM:

Dulbecco’s modified Eagle’s medium

EPA:

eicosapentaenoic acid

FAK:

focal adhesion kinase

PBS:

phosphate-buffered saline

PUFA:

polyunsaturated fatty acid(s)

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Authors and Affiliations

  1. Institute of Food Research, Norwich Research Park, NR4 7UA, Colney, Norwich, UK

    Rosemary G. Clarke, Elizabeth K. Lund, Peter Latham, Andrew C. Pinder & Ian T. Johnson

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  1. Rosemary G. Clarke
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  2. Elizabeth K. Lund
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  3. Peter Latham
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Correspondence to Elizabeth K. Lund.

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Clarke, R.G., Lund, E.K., Latham, P. et al. Effect of eicosapentaenoic acid on the proliferation and incidence of apoptosis in the colorectal cell line HT29. Lipids 34, 1287–1295 (1999). https://doi.org/10.1007/s11745-999-0480-7

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  • DOI: https://doi.org/10.1007/s11745-999-0480-7

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