Tumor Biology

, Volume 35, Issue 10, pp 9717–9723 | Cite as

Genome-wide screen for serum microRNA expression profile in mfat-1 transgenic mice

  • Zhuo Gao
  • Yan Wang
  • Zijian Ren
  • Qiankun Li
  • Ying Wang
  • Yifan Dai
Research Article


n-3 Polyunsaturated fatty acids (n-3 PUFAs) contribute to preventing many types of diseases, including cancer; however, a high n-6 polyunsaturated fatty acids (n-6 PUFAs) intake in modern diets has the opposite effect. Previously, we developed a transgenic mouse model that expresses a gene, fat-1, encoding an n-3 fatty acid desaturase, which converts n-6 PUFAs to n-3 PUFAs in vivo. MicroRNAs (miRNAs) in serum are stable, reproducible, and consistent among individuals of the same species and serve as potential biomarkers for the detection of cancers and other diseases. Employing illumina sequencing, we analyzed all the serum miRNAs in wild-type and mfat-1 transgenic mice. Using quantitative real-time PCR (RT-qPCR), we identified 12 miRNAs that were highly expressed in mfat-1 mice. Pathway analysis of targets regulated by these miRNAs revealed a significant number of genes involved in the development of cancer, including phosphatidylinositol 3-kinase (PI3K), mitogen-activated protein kinases (MAPK), and mammalian target of rapamycin (mTOR), which suggested a relationship between n-3 PUFAs and cancer prevention.


mfat-1 transgenic mice Illumina sequencing miRNA n-3 PUFA Cancer 



This work was supported by a grant from Jiangsu Key Laboratory of Xenotransplantation (BM2012116).

Conflicts of interest


Supplementary material

13277_2014_2261_MOESM1_ESM.jpg (182 kb)
Supplement 1 (A) Partial gas chromatograph traces showing the polyunsaturated fatty-acid profiles of total lipids extracted from different groups of mice. The n-6 PUFA peak is marked in black while the n-3 PUFA peak is in red. The mice in all groups were fed a normal diet. (B) Measurement of n-6/ n-3 PUFA ratio in mouse tails (12-weeks-old) by gas chromatographic analysis. The compositions of n-6 or n-3 PUFAs are expressed using relative percentages; i.e., the distribution areas of n-3 or n-6 PUFA peaks divided by the total peak areas of all detectable saturated and unsaturated free fatty acids (from the same sample) resolved from the gas chromatography column. Values are the means ± SD (n=16) in different groups. (JPEG 181 kb)
13277_2014_2261_MOESM2_ESM.xlsx (39 kb)
Supplement 2 Illumina sequencing data of serum microRNAs (XLSX 39 kb)


  1. 1.
    Simopoulos AP. Overview of evolutionary aspects of omega 3 fatty acids in the diet. World Rev Nutr Diet. 1998;83:1–11.CrossRefPubMedGoogle Scholar
  2. 2.
    Simopoulos AP. Essential fatty acids in health and chronic diseases. Forum Nutr. 2003;56:67–70.PubMedGoogle Scholar
  3. 3.
    Kang JX, Liu A. The role of the tissue omega-6/omega-3 fatty acid ratio in regulating tumor angiogenesis. Cancer Metastasis Rev. 2013;32:201–10.CrossRefPubMedGoogle Scholar
  4. 4.
    Saedisomeolia A, Wood LG, Garg ML, Gibson PG, Wark PA. Anti-inflammatory effects of long-chain n-3 pufa in rhinovirus-infected cultured airway epithelial cells. Br J Nutr. 2009;101:533–40.CrossRefPubMedGoogle Scholar
  5. 5.
    White PJ, Arita M, Taguchi R, Kang JX, Marette A. Transgenic restoration of long-chain n-3 fatty acids in insulin target tissues improves resolution capacity and alleviates obesity-linked inflammation and insulin resistance in high-fat-fed mice. Diabetes. 2010;59:3066–73.PubMedCentralCrossRefPubMedGoogle Scholar
  6. 6.
    Weylandt KH, Krause LF, Gomolka B, et al. Suppressed liver tumorigenesis in fat-1 mice with elevated omega-3 fatty acids is associated with increased omega-3 derived lipid mediators and reduced tnf-alpha. Carcinogenesis. 2011;32:897–903.PubMedCentralCrossRefPubMedGoogle Scholar
  7. 7.
    Boudrault C, Bazinet RP, Ma DW. Experimental models and mechanisms underlying the protective effects of n-3 polyunsaturated fatty acids in Alzheimer’s disease. J Nutr Biochem. 2009;20:1–10.CrossRefPubMedGoogle Scholar
  8. 8.
    Lee SP, Dart AM, Walker KZ, et al. Effect of altering dietary n-6:N-3 pufa ratio on cardiovascular risk measures in patients treated with statins: a pilot study. Br J Nutr. 2012;108:1280–5.CrossRefPubMedGoogle Scholar
  9. 9.
    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.PubMedCentralCrossRefPubMedGoogle Scholar
  10. 10.
    Chen Z, Zhang Y, Jia C, et al. Mtorc1/2 targeted by n-3 polyunsaturated fatty acids in the prevention of mammary tumorigenesis and tumor progression. Oncogene. 2013. doi: 10.1038/onc.2013.402.Google Scholar
  11. 11.
    Spychalla JP, Kinney AJ, Browse J. Identification of an animal omega-3 fatty acid desaturase by heterologous expression in arabidopsis. Proc Natl Acad Sci U S A. 1997;94:1142–7.PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Kang JX, Wang J, Wu L, Kang ZB. Transgenic mice: fat-1 mice convert n-6 to n-3 fatty acids. Nature. 2004;427:504.CrossRefPubMedGoogle Scholar
  13. 13.
    Wei D, Li J, Shen M, et al. Cellular production of n-3 pufas and reduction of n-6-to-n-3 ratios in the pancreatic beta-cells and islets enhance insulin secretion and confer protection against cytokine-induced cell death. Diabetes. 2010;59:471–8.PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Gravaghi C, La Perle KM, Ogrodwski P, et al. Cox-2 expression, pge(2) and cytokines production are inhibited by endogenously synthesized n-3 pufas in inflamed colon of fat-1 mice. J Nutr Biochem. 2011;22:360–5.CrossRefPubMedGoogle Scholar
  15. 15.
    Bellenger J, Bellenger S, Bataille A, et al. High pancreatic n-3 fatty acids prevent stz-induced diabetes in fat-1 mice: inflammatory pathway inhibition. Diabetes. 2011;60:1090–9.PubMedCentralCrossRefPubMedGoogle Scholar
  16. 16.
    Kang JX. From fat to fat-1: a tale of omega-3 fatty acids. J Membr Biol. 2005;206:165–72.CrossRefPubMedGoogle Scholar
  17. 17.
    Wu X, Ouyang H, Duan B, et al. Production of cloned transgenic cow expressing omega-3 fatty acids. Transgenic Res. 2012;21:537–43.CrossRefPubMedGoogle Scholar
  18. 18.
    Lai L, Kang JX, Li R, et al. Generation of cloned transgenic pigs rich in omega-3 fatty acids. Nat Biotechnol. 2006;24:435–6.PubMedCentralCrossRefPubMedGoogle Scholar
  19. 19.
    Chen X, Ba Y, Ma L, et al. Characterization of micrornas in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res. 2008;18:997–1006.CrossRefPubMedGoogle Scholar
  20. 20.
    Lovis P, Roggli E, Laybutt DR, et al. Alterations in microrna expression contribute to fatty acid-induced pancreatic beta-cell dysfunction. Diabetes. 2008;57:2728–36.PubMedCentralCrossRefPubMedGoogle Scholar
  21. 21.
    Davidson LA, Wang N, Shah MS, et al. N-3 polyunsaturated fatty acids modulate carcinogen-directed non-coding microrna signatures in rat colon. Carcinogenesis. 2009;30:2077–84.PubMedCentralCrossRefPubMedGoogle Scholar
  22. 22.
    Mandal CC, Ghosh-Choudhury T, Dey N, Choudhury GG, Ghosh-Choudhury N. Mir-21 is targeted by omega-3 polyunsaturated fatty acid to regulate breast tumor csf-1 expression. Carcinogenesis. 2012;33:1897–908.PubMedCentralCrossRefPubMedGoogle Scholar
  23. 23.
    Chen X, Liang H, Guan D, et al. A combination of let-7d, let-7 g and let-7i serves as a stable reference for normalization of serum micrornas. PLoS One. 2013;8:e79652.PubMedCentralCrossRefPubMedGoogle Scholar
  24. 24.
    Peurala H, Greco D, Heikkinen T, et al. Mir-34a expression has an effect for lower risk of metastasis and associates with expression patterns predicting clinical outcome in breast cancer. PLoS One. 2011;6:e26122.PubMedCentralCrossRefPubMedGoogle Scholar
  25. 25.
    Tili E, Michaille JJ, Wernicke D, et al. Mutator activity induced by microrna-155 (mir-155) links inflammation and cancer. Proc Natl Acad Sci U S A. 2011;108:4908–13.PubMedCentralCrossRefPubMedGoogle Scholar
  26. 26.
    Hsu SH, Wang B, Kota J, et al. Essential metabolic, anti-inflammatory, and anti-tumorigenic functions of mir-122 in liver. J Clin Invest. 2012;122:2871–83.PubMedCentralCrossRefPubMedGoogle Scholar
  27. 27.
    Manerba A, Vizzardi E, Metra M, Dei Cas L. N-3 pufas and cardiovascular disease prevention. Futur Cardiol. 2010;6:343–50.CrossRefGoogle Scholar
  28. 28.
    Stirban A, Nandrean S, Gotting C, Stratmann B, Tschoepe D. Effects of n-3 polyunsaturated fatty acids (pufas) on circulating adiponectin and leptin in subjects with type 2 diabetes mellitus. Horm Metab Res 2013Google Scholar
  29. 29.
    Itariu BK, Zeyda M, Hochbrugger EE, et al. Long-chain n-3 pufas reduce adipose tissue and systemic inflammation in severely obese nondiabetic patients: a randomized controlled trial. Am J Clin Nutr. 2012;96:1137–49.CrossRefPubMedGoogle Scholar
  30. 30.
    van der Meij BS, van Bokhorst-de van der Schueren MA, Langius JA, Brouwer IA, van Leeuwen PA. N-3 pufas in cancer, surgery, and critical care: a systematic review on clinical effects, incorporation, and washout of oral or enteral compared with parenteral supplementation. Am J Clin Nutr. 2011;94:1248–65.CrossRefPubMedGoogle Scholar
  31. 31.
    Cao W, Yang W, Fan R, et al. Mir-34a regulates cisplatin-induce gastric cancer cell death by modulating pi3k/akt/survivin pathway. Tumour Biol. 2014;35:1287–95.CrossRefPubMedGoogle Scholar
  32. 32.
    Chen D, Li Y, Mei Y, et al. Mir-34a regulates mesangial cell proliferation via the PDGFR-beta/Ras-MAPK signaling pathway. Cell Mol Life Sci. 2014. doi: 10.1007/s00018-014-1599-y.Google Scholar
  33. 33.
    Genovese G, Ergun A, Shukla SA, et al. Microrna regulatory network inference identifies mir-34a as a novel regulator of tgf-beta signaling in glioblastoma. Cancer Discov. 2012;2:736–49.PubMedCentralCrossRefPubMedGoogle Scholar
  34. 34.
    Zou Z, Bidu C, Bellenger S, Narce M, Bellenger J. N-3 polyunsaturated fatty acids and her2-positive breast cancer: interest of the fat-1 transgenic mouse model over conventional dietary supplementation. Biochimie. 2014;96:22–7.CrossRefPubMedGoogle Scholar
  35. 35.
    Liu S, Li D, Li Q, et al. Micrornas of bombyx mori identified by solexa sequencing. BMC Genomics. 2010;11:148.PubMedCentralCrossRefPubMedGoogle Scholar
  36. 36.
    Cock PJ, Fields CJ, Goto N, Heuer ML, Rice PM. The sanger fastq file format for sequences with quality scores, and the solexa/illumina fastq variants. Nucleic Acids Res. 2010;38:1767–71.PubMedCentralCrossRefPubMedGoogle Scholar
  37. 37.
    Bitting RL, Armstrong AJ. Targeting the pi3k/akt/mtor pathway in castration-resistant prostate cancer. Endocr Relat Cancer. 2013;20:R83–99.CrossRefPubMedGoogle Scholar
  38. 38.
    Ghayad SE, Cohen PA. Inhibitors of the pi3k/akt/mtor pathway: new hope for breast cancer patients. Recent Pat Anticancer Drug Discov. 2010;5:29–57.CrossRefPubMedGoogle Scholar
  39. 39.
    Yong HY, Koh MS, Moon A. The p38 mapk inhibitors for the treatment of inflammatory diseases and cancer. Expert Opin Investig Drugs. 2009;18:1893–905.CrossRefPubMedGoogle Scholar
  40. 40.
    Coothankandaswamy V, Liu Y, Mao SC, et al. The alternative medicine pawpaw and its acetogenin constituents suppress tumor angiogenesis via the hif-1/vegf pathway. J Nat Prod. 2010;73:956–61.PubMedCentralCrossRefPubMedGoogle Scholar
  41. 41.
    Toya H, Oyama T, Ohwada S, et al. Immunohistochemical expression of the beta-catenin-interacting protein b9l is associated with histological high nuclear grade and immunohistochemical erbb2/her-2 expression in breast cancers. Cancer Sci. 2007;98:484–90.CrossRefPubMedGoogle Scholar
  42. 42.
    Zhang N, Su Y, Xu L. Targeting pkcepsilon by mir-143 regulates cell apoptosis in lung cancer. FEBS Lett. 2013;587:3661–7.CrossRefPubMedGoogle Scholar
  43. 43.
    Liu L, Yu X, Guo X, et al. Mir-143 is downregulated in cervical cancer and promotes apoptosis and inhibits tumor formation by targeting bcl-2. Mol Med Rep. 2012;5:753–60.PubMedGoogle Scholar
  44. 44.
    Liu C, Kelnar K, Liu B, et al. The microrna mir-34a inhibits prostate cancer stem cells and metastasis by directly repressing cd44. Nat Med. 2011;17:211–5.PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  1. 1.State Key Laboratory of Reproductive Medicine, Jiangsu Key Laboratory of XenotransplantationNanjing Medical UniversityNanjingChina

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