Clinical & Experimental Metastasis

, Volume 32, Issue 4, pp 323–333 | Cite as

Dietary soy isoflavones increase metastasis to lungs in an experimental model of breast cancer with bone micro-tumors

  • Xujuan Yang
  • Aashvini Belosay
  • James A. Hartman
  • Huaxin Song
  • Yukun Zhang
  • Wendan Wang
  • Daniel R. Doerge
  • William G. HelferichEmail author
Research Paper


Bone is one of the most common sites for metastasis in breast cancer (BC). Micro-metastasis in bone marrow was detected in 30 % of patients with stage I, II, or III BC at primary surgery and is a strong indicator of poor prognosis. The role dietary soy isoflavones play in BC with bone micro-metastasis is unclear. In this study, we examined the effects of genistein, daidzein, (−)-equol or a mixture of soy isoflavones on BC with bone micro-metastasis using an experimental model of murine mammary cancer 4T1 cells engineered with luciferase. A small number (1000) of 4T1 cells were injected into the tibia of female Balb/c mice to establish micro-tumors in bone. Soy isoflavones were supplemented in the AIN-93G diet at 750 mg/kg and were provided to mice from 3 weeks before to 3 weeks after cell injection. Bioluminescent imaging was conducted on day 2 (D2), D6, D8, D16 and D20 post cell injection and the results indicated dietary soy isoflavones enhanced the growth of bone micro-tumors on D8. Furthermore, dietary soy isoflavones stimulated metastatic tumor formation in lungs and increased Ki-67 protein expression in these metastasized tumors. In vitro, soy isoflavones (<10 µM) had limited effects on the growth, motility or invasion of 4T1 cells. Thus, the in vivo stimulatory effect could be likely due to systemic effects between the host, 4T1 tumors and soy isoflavones. In conclusion, soy isoflavones stimulate BC with bone micro-metastasis in mice and further investigations are needed regarding their consumption by BC survivors.


Dietary soy isoflavones Breast cancer Bone micro-metastasis Lung metastasis 



Bioluminescence imaging


Breast cancer


Hematoxylin and eosin








Mixed isoflavones


Heat-Inactivated Fetal Bovine Serum



This project was made possible by the National Cancer Institute Grant Number [CA77355] and [P50AT006268] to (WGH) from the National Center for Complementary and Alternative Medicines (NCCAM), the Office of Dietary Supplements (ODS) and the National Cancer Institute (NCI). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NCCAM, ODS, NCI or the National Institutes of Health. In addition, the views expressed in this paper do not necessarily reflect those of the U.S. Food and Drug Administration.

Conflict of Interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in the study involving animals were in accordance with the ethical standards of the IACUC of the UIUC.

Supplementary material

10585_2015_9709_MOESM1_ESM.docx (22 kb)
Supplementary material 1 (DOCX 21 kb)
10585_2015_9709_MOESM2_ESM.docx (691 kb)
Supplementary material 2 (DOCX 690 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Xujuan Yang
    • 1
  • Aashvini Belosay
    • 1
  • James A. Hartman
    • 1
  • Huaxin Song
    • 2
  • Yukun Zhang
    • 1
  • Wendan Wang
    • 1
  • Daniel R. Doerge
    • 3
  • William G. Helferich
    • 1
    Email author
  1. 1.Department of Food Science and Human NutritionUniversity of Illinois at Urbana-ChampaignUrbanaUSA
  2. 2.Health Sciences Center, School of NursingTexas Tech UniversityLubbockUSA
  3. 3.Division of Biochemical Toxicology, National Center for Toxicological ResearchU.S. Food and Drug AdministrationJeffersonUSA

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