Abstract
There is experimental evidence that dietary fish oil, which contains the n−3 fatty acid family, i.e., EPA and DHA, protects against colon tumor development, in part by increasing apoptosis. Since mitochondria can act as central executioners of apoptosis, we hypothesized that EPA and DHA incorporation into colonocyte mitochondrial membranes, owing to their high degree of unsaturation, would enhance susceptibility to damage by reactive oxygen species (ROS) generated via oxidative phosphorylation. This, in turn, would compromise mitochondrial function, thereby initiating apoptosis. To test this hypothesis, colonic crypts were isolated from rats fed either fish oil, purified n−3 fatty acid ethyl esters, or corn oil (control). Dietary lipid source had no effect on colonic mitochondrial phospholipid class mole percentages, although incorporation of EPA and DHA was associated with a reduction in n−6 fatty acids known to enhance colon tumor development, i.e., linoleic acid (LNA) and its metabolic product, arachidonic acid (ARA). Select compositional changes in major phospholipid pools were correlated to alterations in mitochondrial function as assessed by confocal microscopy. The mol% sum of LNA plus ARA in cardiolipin was inversely correlated with ROS (P=0.024). Ethanolamine glycerophospholipid ARA (P=0.046) and choline glycerophospholipid INA (P=0.033) levels were positively correlated to mitochondrial membrane potential. In contrast, ethanolamine glycerophospholipid EPA (P=0.042) and DHA (P=0.024), levels were negatively correlated to mitochondrial membrane potential. Additionally, EPA and DHA levels in choline glycerophospholipids (P=0.026) were positively correlated with caspase 3 activity. These data provide evidence in vivo indicating that dietary FPA and DHA induce compositional changes in colonic mitochondrial membrane phospholipids that facilitate appotosis.
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Abbreviations
- ARA:
-
arachidonic acid (20∶4n−6)
- ChoGpl:
-
choline glycerophospholipids
- CMH2-DCFDA:
-
5 (and 6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate, acetylester
- EtnGpl:
-
ethanolamine glycerophospholipids
- FAEE:
-
fatty acid ethyl ester
- LNA:
-
linoleic acid (18∶2n−6)
- Ptd2Gro:
-
cardiolipin
- ROS:
-
Teactive oxygen species
- Δψmt :
-
mitochondrial membrane potential
References
Anti, M., Giancarlo, M., Armelao, F., Bartoli, G.M., Ficaretlli, R., Percesepe, A., De Vitis, I., Maria, G., Sofo, L., Rapaccini, G.L., Gentiloni, N., Piccioni, E., and Miggiano, G. (1992) Effect of ω-3 Fatty Acids on Rectal Mucosal Cell Proliferation in Subjects at Risk for Colon Cancer, Gastroenterology 103, 883–891.
Bartram, H.P., Gostner, A., Scheppach, W., Reddy, B.S., Rao, C.V., Dusel, G., Richter, F., Richter, A., and Kasper, H. (1993) Effects of Fish Oil on Rectal Cell Proliferation, Mucosal Fatty Acids, and Prostaglandin E2 Release in Healthy Subjects, Gastroenterology 105, 1317–1322.
Anti, M., Armelao, F., Marra, G., Percesepe, A., Bartoli, G.M., Palozza, P., Parrella, P., Canetta, C., Gentiloni, N., De Vitis, I., and Gasbarrini, G. (1994) Effects of Different Doses of Fish Oil on Rectal Cell Proliferation in Patients with Sporadic Colonic Adenomas, Gastroenterology 107, 1709–1718.
Caygill, C.P., Charlett, A., and Hill, M.J. (1996) Fat, Fish, Fish Oil and Cancer, Br. J. Cancer 74, 159–164.
Chang, W.C., Chapkin, R.S., and Lupton, J.R. (1998) Fish Oil Blocks Azoxymethane-Induced Tumorigenesis by Increased Cell Differentiation and Apoptosis Rather Than Decreased Cell Proliferation, J. Nutr. 18, 351–357.
Rao, C.V., Hirose, Y., Indranie, C., and Reddy, B.S. (2001) Modulation of Experimental Colon Tumorigenesis by Types and Amounts of Dietary Fatty Acids. Cancer Res. 61, 1927–1933.
Latham, P., Lund, E.K., and Johnson, I.T. (1999) Dietary n−3 PUFA Increases the Apoptotic Response to 1,2-Dimethylhydrazine, Reduces Mitosis, and Suppresses the Induction of Carcinogenesis in the Rat Colon, Carcinogenesis 20, 645–650.
Hong, M.Y., Lupton, J.R., Morris, J.S., Wang, N., Carroll, R.J., Davidson, L.A., Elder, R.H., and Chapkin, R.S. (2000) Dietary Fish Oil Reduces O 6-Methylguanine DNA Adduct Levels in Rat Colon in Part by Increasing Apoptosis During Tumor Initiation, Cancer Epidemiol. Biomarkers Prev. 9, 819–826.
Chang, W.C., Chapkin, R.S., and Lupton, J.R. (1997) Predictive Value of Proliferation, Differentiation and Apoptosis as Intermediate Markers for Colon Tumorigenesis, Carcinogenesis 18, 721–730.
Green, D.R., and Reed, J.C. (1998) Mitochondria and Apoptosis, Science 281, 1309–1312.
Malis, C.D., Weber, P.C., Leaf, A., and Bonventure, J.V. (1990) Incorporation of Marine Lipids into Mitochondrial Membranes Increases Susceptibility to Damage by Calcium and Reactive Oxygen Species: Evidence for Enhanced Activation of Phospholipase A2 in Mitochondria Enriched with n−3 Fatty Acids, Proc. Natl. Acad. Sci. USA 87, 8845–8849.
Watkins, S.M., Carter, L.C., and German, J.B. (1998) Docosahexaenoic Acid Accumulates in Cardiolipin and Enhances HT-29 Cell Oxidant Production, J. Lipid Res. 39, 1583–1588.
Pickering, J.S., Lupton, J.R., and Chapkin, R.S. (1995) Dietary Fat Fiber and Carcinogen Alter Fecal Diacylglycerol Composition and Mass, Cancer Res. 55, 2293–2298.
Zhang, J., Wu, G., Chapkin, R.S., and Lupton, J.R. (1998) Energy Metabolism of Colonocytes Changes During Tumorigenesis Process and Is Dependent on Diet and Carcinogen, J. Nutr. 128, 1262–1269.
Thompson, D.C., Barhoumi, R., and Burghardt, R.C. (1998) Comparative Toxicity of Eugenol and its Quinone Methide Metabolite in Cultured Liver Cells Using Kinetic Fluorescence Bioassays, Toxicol. Appl. Pharmacol. 149, 55–63.
Johnson, D., and Lardy, H. (1967) Isolation of Liver of Kidney Mitochondria, Methods Enzymol. 15, 94–96.
Michell, R.H., and Hawthorne, J.N. (1965) The Site of Diphosphoinositide Synthesis in Rat Liver, Biochem. Biophys. Res. Commun. 21, 333–338.
Duck-Chong, C.G. (1979) A Rapid Sensitive Method for Determining Phospholipid Phosphorus Involving the Digestion with Magnesium, Lipids 14, 492–497.
Smith, L. (1955) Cytochrome c Oxidase Assay, Methods Biochem. Anal. 2, 427–434.
Maunsback, A.B. (1997) Fixation of Cells in Tissues for Transmission Electron Microscopy, in Cell Biology, a Laboratory Handbook (Celis, J.E., ed.) Vol. 2, Academic Press, San Diego, pp. 105–116.
Hanaichi, T., Sato, T., Iwamoto, T., Malavasi-Yamashiro, J., Hoshino, M., and Mizuno, N. (1986) A Stable Lead by Modification of Sato’s Method, J. Electron Microsc. (Tokyo) 35, 304–306.
Chapkin, R.S., and Carmichael, S.L. (1990) Effects of Dietary n−3 and n−6 Polyunsaturated Fatty Acids on Macrophage Phospholipid Classes and Subclasses, Lipids 25, 827–834.
Barhoumi, R., and Burghardt, R.C. (1996) Kinetic Analysis of the Chronology of Patulin- and Gossypol-Induced Cytotoxicity in vitro, Fundam. Appl. Toxicol. 30, 290–297.
Fan, Y.Y., Zhang, J., Barhoumi, R., Burghardt, R.C., Turner, N.D., Lupton, J.R., and Chapkin, R.S. (1999) Antagonism of CD95 Signaling Blocks Butyrate Induction of Apoptosis in Young Adult Mouse Colonic Cells, Am. J. Physiol. 277, C310-C319.
Zoran, D.L., Barhoumi, R., Burghardt, R.C., Chapkin, R.S., and Lupton, J.R. (1997) Wheat Bran Reduces Tumor Incidence in a Rodent Model of Colon Cancer Independent of Effects on Distal Luminal Butyrate Concentrations, Nutr. Cancer 27, 222–230.
Collett, E.D., Davidson, L.A., Fan, Y.Y., Lupton, J.R., and Chapkin, R.S. (2001) n−6 and n−3 Polyunsaturated Fatty Acids Differentially Modulate Oncogenic Ras Activation in Colonocytes, Am. J. Physiol. 280, C1066-C1075.
Paradies, G., Ruggiero, F.M., Petrosillo, G., and Quagliariello, E. (1998) Peroxidative Damage to Cardiac Mitochondria: Cytochrome Oxidase and Cardiolipin Alterations, FEBS Lett 424, 155–158.
Daum, G., and Vance, J.E. (1997) Import of Lipids into Mitochondria, Prog. Lipid Res. 36, 103–130.
Yamaoka, S., Urade, R., and Kito, M. (1988) Mitochondrial Function in Rats Is Affected by Modification of Membrane Phospholipids with Dietary Sardine Oil, J. Nutr. 118, 290–296.
Surette, M.E., Winkler, J.D., Fonteh, A.N., and Chilton, F.H. (1996) Relationship Between Arachidonate-Phospholipid Remodeling and Apoptosis, Biochemistry 35, 9187–9196.
Stillwell, W., Jenski, L., Crump, F.T., and Ehringer, W. (1997) Effect of Docosahexaenoic Acid on Mouse Mitochondrial Membrane Properties, Lipids 32, 497–506.
Chapkin, R.S., Akoh, C.C., and Miller, C.C. (1991) Influence of Dietary n−3 Fatty Acids on Macrophage Glycerophospholipid Molecular Species and Peptido Leukotriene Synthesis, J. Lipid Res. 32, 1205–1213.
Shidoji, Y., Hayashi, K., Komura, S., Ohishi, N., and Yagi, K. (1999) Loss of Molecular Interaction Between Cytochrome c and Cardiolipin Due to Lipid Peroxidation, Biochem. Biophys. Res. Commun. 264, 343–347.
Paltauf, F., and Polheim, D. (1970) Occurrence of C20 Alk-1-enyl and Alkyl Glycerol Ethers in Phospholipids of the Rat Intestinal Mucosa, Biochim. Biophys. Acta 210, 187–189.
Gaposchkin, D.P., and Zoeller, R.A. Plasmalogen Status Influences Docosahexaenoic Acid Levels in a Macrophage Cell Line. Insights Using Ether Lipid-Deficient Variants, J. Lipid Res. 40, 495–503.
Reiss, D., Beyer, K., and Engelmann, B. (1997) Delayed Oxidative Degradation of Polyunsaturated Diacyl Phospholipids in the Presence of Plasmalogen Phospholipids in Vitro, Biochem. J. 323, 807–814.
Kroemer, G., Dallaporta, B., and Resche-Rigon, M. (1998) The Mitochondrial Death/Life Regulator in Apoptosis and Necrosis, Annu. Rev. Physiol. 60, 619–642.
Heerdt, B.G., Houston, M.A., Anthony, G.M., and Augenlicht, L.H. (1998) Mitochondrial Membrane Potential (Δψmt) in the Coordination of p53-Independent Proliferation and Apoptosis Pathways in Human Colonic Carcinoma Cells, Cancer Res. 58, 2869–2875.
Chapkin, R.S., and Lupton, J.R. (1999) Colonic Cell Proliferation and Apoptosis in Rodent Species: Modulation by Diet, Adv. Exp. Med. Biol. 470, 105–118.
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Chapkin, R.S., Young Hong, M., Fan, YY. et al. Dietary n−3 PUFA alter colonocyte mitochondrial membrane composition and function. Lipids 37, 193–199 (2002). https://doi.org/10.1007/s11745-002-0880-8
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DOI: https://doi.org/10.1007/s11745-002-0880-8