Breast Cancer Research and Treatment

, Volume 141, Issue 3, pp 341–352 | Cite as

DHA is a more potent inhibitor of breast cancer metastasis to bone and related osteolysis than EPA

  • Md Mizanur RahmanEmail author
  • Jyothi Maria Veigas
  • Paul J. Williams
  • Gabriel FernandesEmail author
Preclinical Study


Breast cancer patients often develop bone metastasis evidenced by osteolytic lesions, leading to severe pain and bone fracture. Attenuation of breast cancer metastasis to bone and associated osteolysis by fish oil, rich in EPA and DHA, has been demonstrated previously. However, it was not known whether EPA and DHA differentially or similarly affect breast cancer bone metastasis and associated osteolysis. In vitro culture of parental and luciferase gene encoded MDA-MB-231 human breast cancer cell lines treated with EPA and DHA revealed that DHA inhibits proliferation and invasion of breast cancer cells more potently than EPA. Intra-cardiac injection of parental and luciferase gene encoded MDA-MB-231 cells to athymic NCr nu/nu mice demonstrated that DHA-treated mice had significantly less breast cancer cell burden in bone, and also significantly less osteolytic lesions than EPA-treated mice. In vivo cell migration assay as measured by luciferase intensity revealed that DHA attenuated cell migration specifically to the bone. Moreover, the DHA-treated group showed reduced levels of CD44 and TRAP positive area in bone compared to EPA-treated group. Breast cancer cell burden and osteolytic lesions were also examined in intra-tibially breast cancer cell injected mice and found less breast cancer cell growth and associated osteolysis in DHA-treated mice as compared to EPA-treated mice. Finally, doxorubicin-resistant MCF-7 (MCF-7dox) human breast cancer cell line was used to examine if DHA can improve sensitization of MCF-7dox cells to doxorubicin. DHA improved the inhibitory effect of doxorubicin on proliferation and invasion of MCF-7dox cells. Interestingly, drug resistance gene P-gp was also down-regulated in DHA plus doxorubicin-treated cells. In conclusion, DHA attenuates breast cancer bone metastasis and associated osteolysis more potently than EPA, possibly by inhibiting migration of breast cancer cell to the bone as well as by inhibiting osteoclastic bone resorption.


Breast cancer Bone metastasis Omega-3 fatty acids Docosahexaenoic acid Osteolysis 



We acknowledge Emily Molina for her kind review of our manuscript for grammatical corrections. We thank Ocean Nutrition Canada for free supply of FO-EPA and FO-DHA for this study. This study was supported by DOD, BC096459, Elsa U. Pardee Foundation 131884/44096 and NIH AG034233 grants.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Donaldson MS (2004) Nutrition and cancer: a review of the evidence for an anti-cancer diet. Nutr J 3:19PubMedCrossRefGoogle Scholar
  2. 2.
    Berquin IM, Edwards IJ, Chen YQ (2008) Multi-targeted therapy of cancer by omega-3 fatty acids. Cancer Lett 269:363–377PubMedCrossRefGoogle Scholar
  3. 3.
    Fetterman JW Jr, Zdanowicz MM (2009) Therapeutic potential of n-3 polyunsaturated fatty acids in disease. Am J Health Syst Pharm 66:1169–1179PubMedCrossRefGoogle Scholar
  4. 4.
    Bagga D, Anders KH, Wang HJ, Glaspy JA (2002) Long-chain n-3-to-n-6 polyunsaturated fatty acid ratios in breast adipose tissue from women with and without breast cancer. Nutr Cancer 42:180–185PubMedCrossRefGoogle Scholar
  5. 5.
    Lands WE, Hamazaki T, Yamazaki K, Okuyama H, Sakai K, Goto Y, Hubbard VS (1990) Changing dietary patterns. Am J Clin Nutr 51:991–993PubMedGoogle Scholar
  6. 6.
    Schley PD, Jijon HB, Robinson LE, Field CJ (2005) Mechanisms of omega-3 fatty acid-induced growth inhibition in MDA-MB-231 human breast cancer cells. Breast Cancer Res Treat 92:187–195PubMedCrossRefGoogle Scholar
  7. 7.
    Cao W, Ma Z, Rasenick MM, Yeh S, Yu J (2012) N-3 poly-unsaturated fatty acids shift estrogen signaling to inhibit human breast cancer cell growth. PLoS One 7:e52838PubMedCrossRefGoogle Scholar
  8. 8.
    Rovito D, Giordano C, Vizza D, Plastina P, Barone I, Casaburi I, Lanzino M, De Amicis F, Sisci D, Mauro L, Aquila S, Catalano S, Bonofiglio D, Ando S (2013) Omega-3 PUFA ethanolamides DHEA and EPEA induce autophagy through PPARgamma activation in MCF-7 breast cancer cells. J Cell Physiol 228:1314–1322PubMedCrossRefGoogle Scholar
  9. 9.
    Sczaniecka AK, Brasky TM, Lampe JW, Patterson RE, White E (2012) Dietary intake of specific fatty acids and breast cancer risk among postmenopausal women in the VITAL cohort. Nutr Cancer 64:1131–1142PubMedCrossRefGoogle Scholar
  10. 10.
    Biondo PD, Brindley DN, Sawyer MB, Field CJ (2008) The potential for treatment with dietary long-chain polyunsaturated n-3 fatty acids during chemotherapy. J Nutr Biochem 19:787–796PubMedCrossRefGoogle Scholar
  11. 11.
    Mundy GR (2002) Metastasis to bone: causes, consequences and therapeutic opportunities. Nat Rev Cancer 2:584–593PubMedCrossRefGoogle Scholar
  12. 12.
    Nguyen DX, Bos PD, Massague J (2009) Metastasis: from dissemination to organ-specific colonization. Nat Rev Cancer 9:274–284PubMedCrossRefGoogle Scholar
  13. 13.
    Kopp HG, Avecilla ST, Hooper AT, Rafii S (2005) The bone marrow vascular niche: home of HSC differentiation and mobilization. Physiology (Bethesda) 20:349–356CrossRefGoogle Scholar
  14. 14.
    Rahman MM, Bhattacharya A, Banu J, Kang JX, Fernandes G (2009) Endogenous n-3 fatty acids protect ovariectomy induced bone loss by attenuating osteoclastogenesis. J Cell Mol Med 13:1833–1844PubMedCrossRefGoogle Scholar
  15. 15.
    Poulsen RC, Firth EC, Rogers CW, Moughan PJ, Kruger MC (2007) Specific effects of gamma-linolenic, eicosapentaenoic, and docosahexaenoic ethyl esters on bone post-ovariectomy in rats. Calcif Tissue Int 81:459–471PubMedCrossRefGoogle Scholar
  16. 16.
    Kruger MC, Coetzee M, Haag M, Weiler H (2010) Long-chain polyunsaturated fatty acids: selected mechanisms of action on bone. Prog Lipid Res 49:438–449PubMedCrossRefGoogle Scholar
  17. 17.
    Bhattacharya A, Rahman M, Sun D, Fernandes G (2007) Effect of fish oil on bone mineral density in aging C57BL/6 female mice. J Nutr Biochem 18:372–379PubMedCrossRefGoogle Scholar
  18. 18.
    Rahman MM, Bhattacharya A, Fernandes G (2008) Docosahexaenoic acid is more potent inhibitor of osteoclast differentiation in RAW 264.7 cells than eicosapentaenoic acid. J Cell Physiol 214:201–209PubMedCrossRefGoogle Scholar
  19. 19.
    Bhattacharya A, Rahman M, Banu J, Lawrence RA, McGuff HS, Garrett IR, Fischbach M, Fernandes G (2005) Inhibition of osteoporosis in autoimmune disease prone MRL/Mpj-Fas(lpr) mice by N-3 fatty acids. J Am Coll Nutr 24:200–209PubMedCrossRefGoogle Scholar
  20. 20.
    Weiss LA, Barrett-Connor E, von Muhlen D (2005) Ratio of n-6 to n-3 fatty acids and bone mineral density in older adults: the Rancho Bernardo Study. Am J Clin Nutr 81:934–938PubMedGoogle Scholar
  21. 21.
    Kruger G, Huber MC, Bonifer C (1999) The −3.9 kb DNaseI hypersensitive site of the chicken lysozyme locus harbours an enhancer with unusual chromatin reorganizing activity. Gene 236:63–77PubMedCrossRefGoogle Scholar
  22. 22.
    Watkins BA, Shen CL, Allen KG, Seifert MF (1996) Dietary (n-3) and (n-6) polyunsaturates and acetylsalicylic acid alter ex vivo PGE2 biosynthesis, tissue IGF-I levels, and bone morphometry in chicks. J Bone Miner Res 11:1321–1332PubMedCrossRefGoogle Scholar
  23. 23.
    Watkins BA, Li Y, Seifert MF (2006) Dietary ratio of n-6/n-3 PUFAs and docosahexaenoic acid: actions on bone mineral and serum biomarkers in ovariectomized rats. J Nutr Biochem 17:282–289PubMedCrossRefGoogle Scholar
  24. 24.
    Maggio M, Artoni A, Lauretani F, Borghi L, Nouvenne A, Valenti G, Ceda GP (2009) The impact of omega-3 fatty acids on osteoporosis. Curr Pharm Des 15:4157–4164PubMedCrossRefGoogle Scholar
  25. 25.
    Moselhy SS, Al-Malki AL, Kumosani TA, Jalal JA (2012) Modulatory effect of cod liver oil on bone mineralization in overiectomized female Sprague Dawley rats. Toxicol Ind Health 28:387–392PubMedCrossRefGoogle Scholar
  26. 26.
    Kokkinos PP, Shaye R, Alam BS, Alam SQ (1993) Dietary lipids, prostaglandin E2 levels, and tooth movement in alveolar bone of rats. Calcif Tissue Int 53:333–337PubMedCrossRefGoogle Scholar
  27. 27.
    Nawata K, Yamauchi M, Takaoka S, Yamaguchi T, Sugimoto T (2013) Association of n-3 polyunsaturated fatty acid intake with bone mineral density in postmenopausal women. Calcif Tissue Int 93(2):147–54Google Scholar
  28. 28.
    Farina EK, Kiel DP, Roubenoff R, Schaefer EJ, Cupples LA, Tucker KL (2011) Protective effects of fish intake and interactive effects of long-chain polyunsaturated fatty acid intakes on hip bone mineral density in older adults: the Framingham Osteoporosis Study. Am J Clin Nutr 93:1142–1151PubMedCrossRefGoogle Scholar
  29. 29.
    Lage S, Bueno M, Andrade F, Prieto JA, Delgado C, Legarda M, Sanjurjo P, Aldamiz-Echevarria LJ (2010) Fatty acid profile in patients with phenylketonuria and its relationship with bone mineral density. J Inherit Metab Dis 33 Suppl 3:363–71Google Scholar
  30. 30.
    Kruger MC, Schollum LM (2005) Is docosahexaenoic acid more effective than eicosapentaenoic acid for increasing calcium bioavailability? Prostaglandins Leukot Essent Fatty Acids 73:327–334PubMedCrossRefGoogle Scholar
  31. 31.
    Watkins BA, Li Y, Allen KG, Hoffmann WE, Seifert MF (2000) Dietary ratio of (n-6)/(n-3) polyunsaturated fatty acids alters the fatty acid composition of bone compartments and biomarkers of bone formation in rats. J Nutr 130:2274–2284PubMedGoogle Scholar
  32. 32.
    Oliver E, McGillicuddy FC, Harford KA, Reynolds CM, Phillips CM, Ferguson JF, Roche HM (2012) Docosahexaenoic acid attenuates macrophage-induced inflammation and improves insulin sensitivity in adipocytes-specific differential effects between LC n-3 PUFA. J Nutr Biochem 23:1192–1200PubMedCrossRefGoogle Scholar
  33. 33.
    Rahman M, Kundu JK, Shin JW, Na HK, Surh YJ (2011) Docosahexaenoic acid inhibits UVB-induced activation of NF-kappaB and expression of COX-2 and NOX-4 in HR-1 hairless mouse skin by blocking MSK1 signaling. PLoS One 6:e28065PubMedCrossRefGoogle Scholar
  34. 34.
    Martinez-Micaelo N, Gonzalez-Abuin N, Terra X, Richart C, Ardevol A, Pinent M, Blay M (2012) Omega-3 docosahexaenoic acid and procyanidins inhibit cyclo-oxygenase activity and attenuate NF-kappaB activation through a p105/p50 regulatory mechanism in macrophage inflammation. Biochem J 441:653–663PubMedCrossRefGoogle Scholar
  35. 35.
    Yuan J, Akiyama M, Nakahama K, Sato T, Uematsu H, Morita I (2010) The effects of polyunsaturated fatty acids and their metabolites on osteoclastogenesis in vitro. Prostaglandins Other Lipid Mediat 92:85–90PubMedCrossRefGoogle Scholar
  36. 36.
    Li Y, Seifert MF, Lim SY, Salem N Jr, Watkins BA (2013) Bone mineral content is positively correlated to n-3 fatty acids in the femur of growing rats. Br J Nutr 104:674–685CrossRefGoogle Scholar
  37. 37.
    Li Y, Seifert MF, Lim SY, Salem N Jr, Watkins BA (2010) Bone mineral content is positively correlated to n-3 fatty acids in the femur of growing rats. Br J Nutr 104:674–685PubMedCrossRefGoogle Scholar
  38. 38.
    Poulsen RC, Moughan PJ, Kruger MC (2008) Docosahexaenoic acid and 17 beta-estradiol co-treatment is more effective than 17 beta-estradiol alone in maintaining bone post-ovariectomy. Exp Biol Med (Maywood) 233:592–602CrossRefGoogle Scholar
  39. 39.
    Jourdan ML, Maheo K, Barascu A, Goupille C, De Latour MP, Bougnoux P, Rio PG (2007) Increased BRCA1 protein in mammary tumours of rats fed marine omega-3 fatty acids. Oncol Rep 17:713–719PubMedGoogle Scholar
  40. 40.
    Sun H, Berquin IM, Owens RT, O’Flaherty JT, Edwards IJ (2008) Peroxisome proliferator-activated receptor gamma-mediated up-regulation of syndecan-1 by n-3 fatty acids promotes apoptosis of human breast cancer cells. Cancer Res 68:2912–2919PubMedCrossRefGoogle Scholar
  41. 41.
    Lu IF, Hasio AC, Hu MC, Yang FM, Su HM (2010) Docosahexaenoic acid induces proteasome-dependent degradation of estrogen receptor alpha and inhibits the downstream signaling target in MCF-7 breast cancer cells. J Nutr Biochem 21:512–517PubMedCrossRefGoogle Scholar
  42. 42.
    Spencer L, Mann C, Metcalfe M, Webb M, Pollard C, Spencer D, Berry D, Steward W, Dennison A (2009) The effect of omega-3 FAs on tumour angiogenesis and their therapeutic potential. Eur J Cancer 45:2077–2086PubMedCrossRefGoogle Scholar
  43. 43.
    Serini S, Trombino S, Oliva F, Piccioni E, Monego G, Resci F, Boninsegna A, Picci N, Ranelletti FO, Calviello G (2008) Docosahexaenoic acid induces apoptosis in lung cancer cells by increasing MKP-1 and down-regulating p-ERK1/2 and p-p38 expression. Apoptosis 13:1172–1183PubMedCrossRefGoogle Scholar
  44. 44.
    Stillwell W, Shaikh SR, Zerouga M, Siddiqui R, Wassall SR (2005) Docosahexaenoic acid affects cell signaling by altering lipid rafts. Reprod Nutr Dev 45:559–579PubMedCrossRefGoogle Scholar
  45. 45.
    Mandal CC, Ghosh-Choudhury T, Yoneda T, Choudhury GG, Ghosh-Choudhury N (2011) Fish oil prevents breast cancer cell metastasis to bone. Biochem Biophys Res Commun 402:602–607CrossRefGoogle Scholar
  46. 46.
    Hiraga T, Williams PJ, Mundy GR, Yoneda T (2001) The bisphosphonate ibandronate promotes apoptosis in MDA-MB-231 human breast cancer cells in bone metastases. Cancer Res 61:4418–4424PubMedGoogle Scholar
  47. 47.
    Zhang JH, Wang J, Tang J, Barnett B, Dickson J, Hahsimoto N, Williams P, Ma W, Zheng W, Yoneda T, Pageau S, Chen J (2004) Bone sialoprotein promotes bone metastasis of a non-bone-seeking clone of human breast cancer cells. Anticancer Res 24:1361–1368PubMedGoogle Scholar
  48. 48.
    Nagae M, Hiraga T, Wakabayashi H, Wang L, Iwata K, Yoneda T (2006) Osteoclasts play a part in pain due to the inflammation adjacent to bone. Bone 39:1107–1115PubMedCrossRefGoogle Scholar
  49. 49.
    Zhao Y, Bu L, Yan H, Jia W (2009) 20S-protopanaxadiol inhibits P-glycoprotein in multidrug resistant cancer cells. Planta Med 75:1124–1128PubMedCrossRefGoogle Scholar
  50. 50.
    Guise TA, Yin JJ, Taylor SD, Kumagai Y, Dallas M, Boyce BF, Yoneda T, Mundy GR (1996) Evidence for a causal role of parathyroid hormone-related protein in the pathogenesis of human breast cancer-mediated osteolysis. J Clin Invest 98:1544–1549PubMedCrossRefGoogle Scholar
  51. 51.
    Ouhtit A, Abd Elmageed ZY, Abdraboh ME, Lioe TF, Raj MH (2007) In vivo evidence for the role of CD44s in promoting breast cancer metastasis to the liver. Am J Pathol 171:2033–2039PubMedCrossRefGoogle Scholar
  52. 52.
    Weber GF, Bronson RT, Ilagan J, Cantor H, Schmits R, Mak TW (2002) Absence of the CD44 gene prevents sarcoma metastasis. Cancer Res 62:2281–2286PubMedGoogle Scholar
  53. 53.
    Gotte M, Yip GW (2006) Heparanase, hyaluronan, and CD44 in cancers: a breast carcinoma perspective. Cancer Res 66:10233–10237PubMedCrossRefGoogle Scholar
  54. 54.
    Sheridan C, Kishimoto H, Fuchs RK, Mehrotra S, Bhat-Nakshatri P, Turner CH, Goulet R Jr, Badve S, Nakshatri H (2006) CD44+/CD24− breast cancer cells exhibit enhanced invasive properties: an early step necessary for metastasis. Breast Cancer Res 8:R59PubMedCrossRefGoogle Scholar
  55. 55.
    Nakamura H, Hiraga T, Ninomiya T, Hosoya A, Fujisaki N, Yoneda T, Ozawa H (2008) Involvement of cell-cell and cell-matrix interactions in bone destruction induced by metastatic MDA-MB-231 human breast cancer cells in nude mice. J Bone Miner Metab 26:642–647PubMedCrossRefGoogle Scholar
  56. 56.
    Hill A, McFarlane S, Johnston PG, Waugh DJ (2006) The emerging role of CD44 in regulating skeletal micrometastasis. Cancer Lett 237:1–9PubMedCrossRefGoogle Scholar
  57. 57.
    Tumber A, Morgan HM, Meikle MC, Hill PA (2001) Human breast-cancer cells stimulate the fusion, migration and resorptive activity of osteoclasts in bone explants. Int J Cancer 91:665–672PubMedCrossRefGoogle Scholar
  58. 58.
    Chen HW, Chao CY, Lin LL, Lu CY, Liu KL, Lii CK, Li CC (2013) Inhibition of matrix metalloproteinase-9 expression by docosahexaenoic acid mediated by heme oxygenase 1 in 12-O-tetradecanoylphorbol-13-acetate-induced MCF-7 human breast cancer cells. Arch Toxicol 87:857–869PubMedCrossRefGoogle Scholar
  59. 59.
    Yamagami T, Porada CD, Pardini RS, Zanjani ED, Almeida-Porada G (2009) Docosahexaenoic acid induces dose dependent cell death in an early undifferentiated subtype of acute myeloid leukemia cell line. Cancer Biol Ther 8:331–337PubMedCrossRefGoogle Scholar
  60. 60.
    Menendez JA, Lupu R, Colomer R (2005) Exogenous supplementation with omega-3 polyunsaturated fatty acid docosahexaenoic acid (DHA; 22:6n-3) synergistically enhances taxane cytotoxicity and downregulates Her-2/neu (c-erbB-2) oncogene expression in human breast cancer cells. Eur J Cancer Prev 14:263–270PubMedCrossRefGoogle Scholar
  61. 61.
    Baumgartner M, Sturlan S, Roth E, Wessner B, Bachleitner-Hofmann T (2004) Enhancement of arsenic trioxide-mediated apoptosis using docosahexaenoic acid in arsenic trioxide-resistant solid tumor cells. Int J Cancer 112:707–712PubMedCrossRefGoogle Scholar
  62. 62.
    Bougnoux P, Hajjaji N, Ferrasson MN, Giraudeau B, Couet C, Le Floch O (2009) Improving outcome of chemotherapy of metastatic breast cancer by docosahexaenoic acid: a phase II trial. Br J Cancer 101:1978–1985PubMedCrossRefGoogle Scholar
  63. 63.
    Maheo K, Vibet S, Steghens JP, Dartigeas C, Lehman M, Bougnoux P, Gore J (2005) Differential sensitization of cancer cells to doxorubicin by DHA: a role for lipoperoxidation. Free Radic Biol Med 39:742–751PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  1. 1.Department of MedicineUniversity of Texas Health Science Center at San AntonioSan AntonioUSA

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