Fish oil supplementation induces expression of genes related to cell cycle, endoplasmic reticulum stress and apoptosis in peripheral blood mononuclear cells: a transcriptomic approach

M C W Myhrstad, S M Ulven, C-C Günther, I Ottestad, M Holden, E Ryeng, G I Borge, A Kohler, K W Brønner, M Thoresen, K B Holven, M C W Myhrstad, S M Ulven, C-C Günther, I Ottestad, M Holden, E Ryeng, G I Borge, A Kohler, K W Brønner, M Thoresen, K B Holven

Abstract

Background: Fish oil supplementation has been shown to alter gene expression of mononuclear cells both in vitro and in vivo. However, little is known about the total transcriptome profile in healthy subjects after intake of fish oil. We therefore investigated the gene expression profile in peripheral blood mononuclear cells (PBMCs) after intake of fish oil for 7 weeks using transcriptome analyses.

Design: In a 7-week, double-blinded, randomized, controlled, parallel-group study, healthy subjects received 8 g day(-1) fish oil (1.6 g day(-1) eicosapentaenoic acid + docosahexaenoic acid) (n = 17) or 8 g day(-1) high oleic sunflower oil (n = 19). Microarray analyses of RNA isolated from PBMCs were performed at baseline and after 7 weeks of intervention.

Results: Cell cycle, DNA packaging and chromosome organization are biological processes found to be upregulated after intake of fish oil compared to high oleic sunflower oil using a moderated t-test. In addition, gene set enrichment analysis identified several enriched gene sets after intake of fish oil. The genes contributing to the significantly different gene sets in the subjects given fish oil compared with the control group are involved in cell cycle, endoplasmic reticulum (ER) stress and apoptosis. Gene transcripts with common motifs for 35 known transcription factors including E2F, TP53 and ATF4 were upregulated after intake of fish oil.

Conclusion: We have shown that intake of fish oil for 7 weeks modulates gene expression in PBMCs of healthy subjects. The increased expression of genes related to cell cycle, ER stress and apoptosis suggests that intake of fish oil may modulate basic cellular processes involved in normal cellular function.

Trial registration: ClinicalTrials.gov NCT01034423.

Keywords: fish oil; intervention; n-3 fatty acid; peripheral blood mononuclear cells; transcriptome.

© 2014 The Authors. Journal of Internal Medicine published by John Wiley & Sons Ltd on behalf of The Association for the Publication of the Journal of Internal Medicine.

Figures

Fig 1
Fig 1
Flow chart showing subjects enrolled, lost during follow-up, and included in the statistical analysis at baseline and after 7 weeks of fish oil supplementation. FO, fish oil; HOSO, high oleic sunflower oil; oxFO, oxidized fish oil. The oxFO group was not included in the present study.
Fig 2
Fig 2
Metacore analysis of leading edge genes. The pathway map showing the role of APC in cell cycle regulation, identified with the Metacore (GeneGo) pathway tool, among the leading edge genes from gene set enrichment analyses using the gene set collection C2 cgp. The leading edge genes were found to be upregulated after intake of fish oil (FO) compared with high oleic sunflower oil (HOSO). The experimental data are shown on the maps as ‘thermometer-like’ figures. Upward thermometers (red) indicate upregulated gene transcripts in the FO group and downward thermometers (blue) indicate downregulated expression levels of the genes in the HOSO group. Further explanations are provided at http://pathwaymaps.com/pdf/MC_legend.pdf. cgp, chemical and genetic perturbations. APC, anaphase-promoting complex.
Fig 3
Fig 3
Metacore analysis of leading edge genes. The pathway map: apoptosis and endoplasmic reticulum stress, identified with the Metacore (GeneGo) pathway tool among the leading edge genes from gene set enrichment analyses using the gene set collection C2 cgp. The leading edge genes were found to be upregulated after intake of fish oil (FO) compared with high oleic sunflower oil (HOSO). The experimental data are shown on the maps as ‘thermometer-like’ figures. Upward thermometers (red) indicate upregulated gene transcripts in the FO group and downward thermometers (blue) indicate the downregulated expression levels of the genes in the HOSO group. Further explanations are provided at http://pathwaymaps.com/pdf/MC_legend.pdf.

References

    1. Hu FB, Bronner L, Willett WC, et al. Fish and omega-3 fatty acid intake and risk of coronary heart disease in women. JAMA. 2002;287:1815–21.
    1. He K, Song Y, Daviglus ML, et al. Accumulated evidence on fish consumption and coronary heart disease mortality: a meta-analysis of cohort studies. Circulation. 2004;109:2705–11.
    1. GISSI-Prevenzione Investigators. Dietary supplementation with n-3 polyunsaturated fatty acids and vitamin E after myocardial infarction: results of the GISSI-Prevenzione trial. Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto miocardico. Lancet. 1999;354:447–55.
    1. Rizos EC, Ntzani EE, Bika E, Kostapanos MS, Elisaf MS. Association between omega-3 fatty acid supplementation and risk of major cardiovascular disease events: a systematic review and meta-analysis. JAMA. 2012;308:1024–33.
    1. Packard RR, Libby P. Inflammation in atherosclerosis: from vascular biology to biomarker discovery and risk prediction. Clin Chem. 2008;54:24–38.
    1. Bryan S, Baregzay B, Spicer D, Singal PK, Khaper N. Redox-inflammatory synergy in the metabolic syndrome. Can J Physiol Pharmacol. 2013;91:22–30.
    1. Federico A, Morgillo F, Tuccillo C, Ciardiello F, Loguercio C. Chronic inflammation and oxidative stress in human carcinogenesis. Int J Cancer. 2007;121:2381–6.
    1. Calder PC. n-3 polyunsaturated fatty acids, inflammation, and inflammatory diseases. Am J Clin Nutr. 2006;83:1505S–19S.
    1. Lopez-Garcia E, Schulze MB, Manson JE, et al. Consumption of (n-3) fatty acids is related to plasma biomarkers of inflammation and endothelial activation in women. J Nutr. 2004;134:1806–11.
    1. Madsen T, Skou HA, Hansen VE, et al. C-reactive protein, dietary n-3 fatty acids, and the extent of coronary artery disease. Am J Cardiol. 2001;88:1139–42.
    1. Myhrstad MC, Retterstol K, Telle-Hansen VH, et al. Effect of marine n-3 fatty acids on circulating inflammatory markers in healthy subjects and subjects with cardiovascular risk factors. Inflamm Res. 2011;60:309–19.
    1. Kliewer SA, Sundseth SS, Jones SA, et al. Fatty acids and eicosanoids regulate gene expression through direct interactions with peroxisome proliferator-activated receptors alpha and gamma. Proc Natl Acad Sci U S A. 1997;94:4318–23.
    1. Tontonoz P, Spiegelman BM. Fat and beyond: the diverse biology of PPARgamma. Annu Rev Biochem. 2008;77:289–312.
    1. Castrillo A, Tontonoz P. Nuclear receptors in macrophage biology: at the crossroads of lipid metabolism and inflammation. Annu Rev Cell Dev Biol. 2004;20:455–80.
    1. Lee JY, Plakidas A, Lee WH, et al. Differential modulation of Toll-like receptors by fatty acids: preferential inhibition by n-3 polyunsaturated fatty acids. J Lipid Res. 2003;44:479–86.
    1. Oh DY, Talukdar S, Bae EJ, et al. GPR120 is an omega-3 fatty acid receptor mediating potent anti-inflammatory and insulin-sensitizing effects. Cell. 2010;142:687–98.
    1. Ma J, Dempsey AA, Stamatiou D, Marshall KW, Liew CC. Identifying leukocyte gene expression patterns associated with plasma lipid levels in human subjects. Atherosclerosis. 2007;191:63–72.
    1. de Mello VD, Kolehmanien M, Schwab U, Pulkkinen L, Uusitupa M. Gene expression of peripheral blood mononuclear cells as a tool in dietary intervention studies: what do we know so far? Mol Nutr Food Res. 2012;56:1160–72.
    1. Bouwens M, Afman LA, Muller M. Fasting induces changes in peripheral blood mononuclear cell gene expression profiles related to increases in fatty acid beta-oxidation: functional role of peroxisome proliferator activated receptor alpha in human peripheral blood mononuclear cells. Am J Clin Nutr. 2007;86:1515–23.
    1. Bakker GC, van Erk MJ, Pellis L, et al. An antiinflammatory dietary mix modulates inflammation and oxidative and metabolic stress in overweight men: a nutrigenomics approach. Am J Clin Nutr. 2010;91:1044–59.
    1. Rudkowska I, Raymond C, Ponton A, et al. Validation of the use of peripheral blood mononuclear cells as surrogate model for skeletal muscle tissue in nutrigenomic studies. OMICS. 2011;15:1–7.
    1. Ottestad I, Vogt G, Retterstol K, et al. Oxidised fish oil does not influence established markers of oxidative stress in healthy human subjects: a randomised controlled trial. Br J Nutr. 2011;108:315–26.
    1. Ottestad I, Retterstol K, Myhrstad MC, et al. Intake of oxidised fish oil does not affect circulating levels of oxidised LDL or inflammatory markers in healthy subjects. Nutr Metab Cardiovasc Dis. 2013;23:e3–4.
    1. Ottestad I, Hassani S, Borge GI, et al. Fish oil supplementation alters the plasma lipidomic profile and increases long-chain PUFAs of phospholipids and triglycerides in healthy subjects. PLoS One. 2012;7:e42550.
    1. Smyth GK. Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol. 2004;3 Article3.
    1. Brazma A, Hingamp P, Quackenbush J, et al. Minimum information about a microarray experiment (MIAME)-toward standards for microarray data. Nat Genet. 2001;29:365–71.
    1. Vermeulen K, Van Bockstaele DR, Berneman ZN. The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif. 2003;36:131–49.
    1. Bouwens M, van de Rest O, Dellschaft N, et al. Fish-oil supplementation induces antiinflammatory gene expression profiles in human blood mononuclear cells. Am J Clin Nutr. 2009;90:415–24.
    1. Afman LA, Muller M. Human nutrigenomics of gene regulation by dietary fatty acids. Prog Lipid Res. 2012;51:63–70.
    1. Finstad HS, Myhrstad MC, Heimli H, et al. Multiplication and death-type of leukemia cell lines exposed to very long-chain polyunsaturated fatty acids. Leukemia. 1998;12:921–9.
    1. Jakobsen CH, Storvold GL, Bremseth H, et al. DHA induces ER stress and growth arrest in human colon cancer cells: associations with cholesterol and calcium homeostasis. J Lipid Res. 2008;49:2089–100.
    1. Danbara N, Yuri T, Tsujita-Kyutoku M, et al. Conjugated docosahexaenoic acid is a potent inducer of cell cycle arrest and apoptosis and inhibits growth of colo 201 human colon cancer cells. Nutr Cancer. 2004;50:71–9.
    1. Slagsvold JE, Pettersen CH, Storvold GL, Follestad T, Krokan HE, Schonberg SA. DHA alters expression of target proteins of cancer therapy in chemotherapy resistant SW620 colon cancer cells. Nutr Cancer. 2010;62:611–21.
    1. Schroder M, Kaufman RJ. ER stress and the unfolded protein response. Mutat Res. 2005;569:29–63.
    1. Benbrook DM, Long A. Integration of autophagy, proteasomal degradation, unfolded protein response and apoptosis. Exp Oncol. 2012;34:286–97.
    1. Brown MK, Naidoo N. The endoplasmic reticulum stress response in aging and age-related diseases. Front Physiol. 2012;3:263.
    1. Minamino T, Kitakaze M. ER stress in cardiovascular disease. J Mol Cell Cardiol. 2010;48:1105–10.
    1. Zhang K, Kaufman RJ. The unfolded protein response: a stress signaling pathway critical for health and disease. Neurology. 2006;66:S102–9.
    1. van Laar T, Schouten T, Hoogervorst E, van Eck M, van der Eb AJ, Terleth C. The novel MMS-inducible gene Mif1/KIAA0025 is a target of the unfolded protein response pathway. FEBS Lett. 2000;469:123–31.
    1. Olsen KS, Fenton C, Froyland L, Waaseth M, Paulssen RH, Lund E. Plasma Fatty Acid ratios affect blood gene expression profiles – a cross-sectional study of the norwegian women and cancer post-genome cohort. PLoS One. 2013;8:e67270.
    1. Yang Z, Klionsky DJ. Mammalian autophagy: core molecular machinery and signaling regulation. Curr Opin Cell Biol. 2010;22:124–31.
    1. Singh R, Kaushik S, Wang Y, et al. Autophagy regulates lipid metabolism. Nature. 2009;458:1131–5.
    1. Huang JH, Hood DA. Age-associated mitochondrial dysfunction in skeletal muscle: contributing factors and suggestions for long-term interventions. IUBMB Life. 2009;61:201–14.
    1. Gibson GE, Karuppagounder SS, Shi Q. Oxidant-induced changes in mitochondria and calcium dynamics in the pathophysiology of Alzheimer's disease. Ann N Y Acad Sci. 2008;1147:221–32.
    1. Lee CK, Klopp RG, Weindruch R, Prolla TA. Gene expression profile of aging and its retardation by caloric restriction. Science. 1999;285:1390–3.
    1. Nisoli E, Tonello C, Cardile A, et al. Calorie restriction promotes mitochondrial biogenesis by inducing the expression of eNOS. Science. 2005;310:314–7.
    1. Bøhn SK, Myhrstad MC, Thoresen M, et al. Blood cell gene expression associated with cellular stress defense is modulated by antioxidant-rich food in a randomised controlled clinical trial of male smokers. BMC Med. 2010;8:54.
    1. Semenza GL. Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 2003;3:721–32.
    1. Schmidt S, Stahl F, Mutz KO, Scheper T, Hahn A, Schuchardt JP. Different gene expression profiles in normo- and dyslipidemic men after fish oil supplementation: results from a randomized controlled trial. Lipids Health Dis. 2012;11:105.
    1. Schmidt S, Stahl F, Mutz KO, Scheper T, Hahn A, Schuchardt JP. Transcriptome-based identification of antioxidative gene expression after fish oil supplementation in normo- and dyslipidemic men. Nutr Metab. 2012;9:45.
    1. Schmidt S, Willers J, Stahl F, et al. Regulation of lipid metabolism-related gene expression in whole blood cells of normo- and dyslipidemic men after fish oil supplementation. Lipids Health Dis. 2012;11:172.
    1. Rudkowska I, Paradis AM, Thifault E, et al. Transcriptomic and metabolomic signatures of an n-3 polyunsaturated fatty acids supplementation in a normolipidemic/normocholesterolemic Caucasian population. J Nutr Biochem. 2013;24:54–61.
    1. Vedin I, Cederholm T, Freund-Levi Y, et al. Effects of DHA-rich n-3 fatty acidsupplementation on gene expression in blood mononuclear leukocytes: the OmegAD study. PLoS One. 2012;7:e35425.
    1. Afman L, Muller M. Nutrigenomics: from molecular nutrition to prevention of disease. J Am Diet Assoc. 2006;106:569–76.

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