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Bioengineered tumor microenvironments with naked mole rats high-molecular-weight hyaluronan induces apoptosis in breast cancer cells

Oncogene (2019) | Download Citation


The naked mole rat (nmr) is cancer resistant due to the abundant production of extremely high-molecular-weight hyaluronan (EHMW-HA). However, whether EHMW-HA has similar anti-cancer effects in mice and humans remains to be determined. The present study used breast cancer cells to clarify the effect of EHMW-HA on breast cancer. First, the overexpression of nmrHas2 in 4T1 and BT549 cell lines in both two-dimensional (2D) and three-dimensional (3D) models to mimic tumor microenvironment was established. The 4T1/BT549-nmrHas2 cells could secrete EHMW-HA (with a molecular weight of up to 6 MDa), which was similar to that found in the naked mole rat. Second, EHMW-HA altering tumor microenvironment in both 2D monolayers and 3D spheroids significantly enhanced apoptosis, inhibiting the proliferation of 4T1 and BT549 cells. The prominent anticancer effects of EHMW-HA on the cancer-cell apoptosis phenotype were further confirmed by inhibiting tumor formation in nude mice. Finally, EHMW-HA significantly induced higher p53 protein expression, which enhanced pro-apoptotic proteins p21 and Bax in breast cancer cells; this is in contrast with the triggering of hypersensitivity of the naked mole rat cells to early contact inhibition (ECI). These results have important implications for the design of therapeutic approaches based on the application of EHMW-HA.

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  1. 1.

    Karbownik MS, Nowak JZ. Hyaluronan: towards novel anti-cancer therapeutics. Pharmacol Rep. 2013;65:1056–74.

  2. 2.

    Kultti A, Li X, Jiang P, Thompson CB, Frost GI, Shepard HM. Therapeutic targeting of hyaluronan in the tumor stroma. Cancers. 2012;4:873–903.

  3. 3.

    Tian X, Azpurua J, Hine C, Vaidya A, Myakishev-Rempel M, Ablaeva J, et al. High-molecular-mass hyaluronan mediates the cancer resistance of the naked mole rat. Nature. 2013;499:346–9.

  4. 4.

    Tian X, Azpurua J, Ke Z, Augereau A, Zhang ZD, Vijg J, et al. INK4 locus of the tumor-resistant rodent, the naked mole rat, expresses a functional p15/p16 hybrid isoform. Proc Natl Acad Sci USA. 2015;112:1053–8.

  5. 5.

    Heldin P, Basu K, Olofsson B, Porsch H, Kozlova I, Kahata K. Deregulation of hyaluronan synthesis, degradation and binding promotes breast cancer. J Biochem. 2013;154:395–408.

  6. 6.

    Thanos CD. Targeting the physicochemical, cellular, and immunosuppressive properties of the tumor microenvironment by depletion of hyaluronan to treat cancer. In: Novel immunotherapeutic approaches to the treatment of cancer. New York: Springer, Cham, 2016. p. 249–68.

  7. 7.

    Bernert B, Porsch H, Heldin P. Hyaluronan synthase 2 (HAS2) promotes breast cancer cell invasion by suppression of tissue metalloproteinase inhibitor 1 (TIMP-1). J Biol Chem. 2011;286:42349–59.

  8. 8.

    Okuda H, Kobayashi A, Xia B, Watabe M, Pai SK, Hirota S, et al. Hyaluronan synthase HAS2 promotes tumor progression in bone by stimulating the interaction of breast cancer stem-like cells with macrophages and stromal cells. Cancer Res. 2012;72:537–47.

  9. 9.

    Huang Z, Zhao C, Radi A. Characterization of hyaluronan, hyaluronidase PH20, and HA synthase HAS2 in inflammation and cancer. Inflamm Cell Signal. 2014;1.

  10. 10.

    Wu M, Cao M, He Y, Liu Y, Yang C, Du Y, et al. A novel role of low molecular weight hyaluronan in breast cancer metastasis. FASEB J. 2015;29:1290–8.

  11. 11.

    Miyawaki S, Kawamura Y, Oiwa Y, Shimizu A, Hachiya T, Bono H, et al. Tumour resistance in induced pluripotent stem cells derived from naked mole-rats. Nat Commun. 2016;7:11471.

  12. 12.

    Park TJ, Reznick J, Peterson BL, Blass G, Omerbasic D, Bennett NC, et al. Fructose-driven glycolysis supports anoxia resistance in the naked mole-rat. Science. 2017;356:307–11.

  13. 13.

    Carvalho MP, Costa EC, Miguel SP, Correia IJ. Tumor spheroid assembly on hyaluronic acid-based structures: a review. Carbohydr Polym. 2016;150:139–48.

  14. 14.

    Pampaloni F, Reynaud EG, Stelzer EH. The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol. 2007;8:839–45.

  15. 15.

    Gilli R, Kacurakova M, Mathlouthi M, Navarini L, Paoletti S. FTIR studies of sodium hyaluronate and its oligomers in the amorphous solid phase and in aqueous solution. Carbohydr Res. 1994;263:315–26.

  16. 16.

    Newmeyer DD, Ferguson-Miller S. Mitochondria: releasing power for life and unleashing the machineries of death. Cell. 2003;112:481–90.

  17. 17.

    Giorgio M, Migliaccio E, Orsini F, Paolucci D, Moroni M, Contursi C, et al. Electron transfer between cytochrome c and p66Shc generates reactive oxygen species that trigger mitochondrial apoptosis. Cell. 2005;122:221–33.

  18. 18.

    Lee YJ, Kim SA, Lee SH. Hyaluronan suppresses lidocaine-induced apoptosis of human chondrocytes in vitro by inhibiting the p53-dependent mitochondrial apoptotic pathway. Acta Pharmacol Sin. 2016;37:664–73.

  19. 19.

    Zhao YF, Qiao SP, Shi SL, Yao LF, Hou XL, Li CF, et al. Modulating three-dimensional microenvironment with hyaluronan of different molecular weights alters breast cancer cell invasion behavior. ACS Appl Mater Interfaces. 2017;9:9327–38.

  20. 20.

    Song YK, Billiar TR, Lee YJ. Role of galectin-3 in breast cancer metastasis: involvement of nitric oxide. Am J Pathol. 2002;160:1069–75.

  21. 21.

    Nikitovic D, Kouvidi K, Karamanos NK, Tzanakakis GN. The roles of hyaluronan/RHAMM/CD44 and their respective interactions along the insidious pathways of fibrosarcoma progression. Biomed Res Int. 2013;2013:929531.

  22. 22.

    Cooper EH, Forbes MA. Serum hyaluronic acid levels in cancer. Br J Cancer. 1988;58:668.

  23. 23.

    Laurent TC, Fraser JR. Hyaluronan. FASEB J. 1992;6:2397–404.

  24. 24.

    Laurent TC, Laurent UB, Fraser JR. Functions of hyaluronan. Ann Rheum Dis. 1995;54:429.

  25. 25.

    Yang C, Cao M, Liu H, He Y, Xu J, Du Y, et al. The high and low molecular weight forms of hyaluronan have distinct effects on CD44 clustering. J Biol Chem. 2012;287:43094–107.

  26. 26.

    Fuchs K, Hippe A, Schmaus A, Homey B, Sleeman JP, Orian-Rousseau V. Opposing effects of high- and low-molecular weight hyaluronan on CXCL12-induced CXCR4 signaling depend on CD44. Cell Death Dis. 2013;4:e819.

  27. 27.

    Beasley KL, Weiss MA, Weiss RA. Hyaluronic acid fillers: a comprehensive review. Facial Plast Surg. 2009;25:86–94.

  28. 28.

    Rayahin JE, Buhrman JS, Zhang Y, Koh TJ, Gemeinhart RA. High and low molecular weight hyaluronic acid differentially influence macrophage activation. ACS Biomater Sci Eng. 2015;1:481–93.

  29. 29.

    Gao F, Liu Y, He Y, Yang C, Wang Y, Shi X, et al. Hyaluronan oligosaccharides promote excisional wound healing through enhanced angiogenesis. Matrix Biol. 2010;29:107–16.

  30. 30.

    Gao F, Yang CX, Mo W, Liu YW, He YQ. Hyaluronan oligosaccharides are potential stimulators to angiogenesis via RHAMM mediated signal pathway in wound healing. Clin Invest Med Med Clin Et Exp. 2008;31:E106–16.

  31. 31.

    Brown TJ. The development of hyaluronan as a drug transporter and excipient for chemotherapeutic drugs. Curr Pharm Biotechnol. 2008;9:253–60.

  32. 32.

    Udabage L, Brownlee GR, Nilsson SK, Brown TJ. The over-expression of HAS2, Hyal-2 and CD44 is implicated in the invasiveness of breast cancer. Exp Cell Res. 2005;310:205–17.

  33. 33.

    Masters KS, Shah DN, Leinwand LA, Anseth KS. Crosslinked hyaluronan scaffolds as a biologically active carrier for valvular interstitial cells. Biomaterials. 2005;26:2517–25.

  34. 34.

    Rowley JA, Madlambayan G, Mooney DJ. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials. 1999;20:45–53.

  35. 35.

    Goldsworthy TL, Conolly RB, Fransson-Steen R. Apoptosis and cancer risk assessment. Mutat Res. 1996;365:71–90.

  36. 36.

    Louderbough JM, Schroeder JA. Understanding the dual nature of CD44 in breast cancer progression. Mol Cancer Res. 2011;9:1573–86.

  37. 37.

    Levine AJ. p53, the cellular gatekeeper for growth and division. Cell. 1997;88:323–31.

  38. 38.

    Giaccia AJ, Kastan MB. The complexity of p53 modulation: emerging patterns from divergent signals. Genes Dev. 1998;12:2973–83.

  39. 39.

    Godar S, Ince TA, Bell GW, Feldser D, Donaher JL, Bergh J, et al. Growth-inhibitory and tumor-suppressive functions of p53 depend on its repression of CD44 expression. Cell. 2008;134:62–73.

  40. 40.

    Mirzayans R, Andrais B, Scott A, Murray D. New insights into p53 signaling and cancer cell response to DNA damage: implications for cancer therapy. J Biomed Biotechnol. 2012;2012:170325.

  41. 41.

    Seluanov A, Hine C, Azpurua J, Feigenson M, Bozzella M, Mao Z, et al. Hypersensitivity to contact inhibition provides a clue to cancer resistance of naked mole-rat. Proc Natl Acad Sci USA. 2009;106:19352–7.

  42. 42.

    Zhao Y, Tyshkovskiy A, Muñoz-Espín D, Tian X, Serrano M, Magalhaes JP, et al. Naked mole rats can undergo developmental, oncogene-induced and DNA damage-induced cellular senescence. Proc Natl Acad Sci USA. 2018;115:1801.

  43. 43.

    Seluanov A, Gladyshev VN, Vijg J, Gorbunova V. Mechanisms of cancer resistance in long-lived mammals. Nat Rev Cancer. 2018;18:433–41.

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This research was supported by the National Natural Science Foundation of China (Grant No.: 51773050 and 81770923), the Opening Foundation of the State Key Laboratory of Cancer Biology (CBSKL201106), and the author was supported by the Heilongjiang Postdoctoral Fund (No. LBH-Z18068) and a general financial grant from the China Postdoctoral Science Foundation (No. 2018M641837).

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Author notes

  1. These authors contributed equally: Yufang Zhao, Shupei Qiao


  1. School of Life Science and Technology, Harbin Institute of Technology, 150080, Harbin, China

    • Yufang Zhao
    • , Shupei Qiao
    • , Xiaolu Hou
    • , Hui Tian
    • , Shuai Deng
    • , Kangruo Ye
    •  & Weiming Tian
  2. Research Center of Basic Space Science, Space Environment Stimulation and Research Interface, Harbin Institute of Technology, 150080, Harbin, China

    • Yufang Zhao
  3. State Key Laboratory of Cancer Biology and Xijing Hospital of Digestive Diseases, The Fourth Military Medical University, 150080, Harbin, China

    • Yongzhan Nie
  4. Department of Mechanical Engineering, University of Saskatchewan, Saskatoon, SK S7N 5A2, Canada

    • Xiongbiao Chen
  5. Division of Glycoscience, Department of Chemistry, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTHRoyal Institute of Technology, AlbaNova University Center, 106 91, Stockholm, Sweden

    • Hongji Yan


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Correspondence to Weiming Tian.

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