Cancer Chemotherapy and Pharmacology

, Volume 82, Issue 2, pp 185–197 | Cite as

The natural agent 4-vinylphenol targets metastasis and stemness features in breast cancer stem-like cells

  • Hoi-Wing Leung
  • Chun-Hay Ko
  • Grace Gar-Lee Yue
  • Ingrid Herr
  • Clara Bik-San Lau
Original Article


Background and purpose

Cancer stem-like cells (CSC) are regarded as the source of tumour origins, metastasis and drug resistance, and limits current treatment regimens. Previously, we reported the first study of the anti-angiogenic and anti-tumour activities of 4-vinylphenol. To further examine the therapeutic role of 4-vinylphenol, the inhibitory effects of 4-vinylphenol on cancer stemness, drug resistance and metastasis in breast cancer were investigated in the present study.

Study design and methods

We enriched parental MDA-MB-231 cells with CSCs in serum-free medium to give spheroids. The effects of 4-vinylphenol on cancer stemness, metastasis and drug resistance in CSC-enriched MDA-MB-231 cells were studied in vitro and in vivo.


CSC-enriched MDA-MB-231 cells demonstrated higher tumorigenic and metastatic potential. 4-Vinylphenol reduced spheroid formation and ALDH1 expression in CSC-enriched cultures, revealing its inhibitory effects on the traits of CSCs. 4-Vinylphenol suppressed colony formation and cell proliferation. 4VP also inhibited in vitro invasion and in vivo metastasis in zebrafish model. Our results showed that it reduced vimentin expression, suppressed cell migration, affected the expression and/or activity of MMPs, TIMPs and uPA. In addition, the expressions of caspases 3 and 9 were increased upon its treatment, and surprisingly, prolonged treatment did not confer cancer cells with drug resistance to 4-vinylphenol. 4-Vinylphenol probably exhibited its anti-cancer activities via beta-catenin, EGFR and AKT signaling pathways.


4-Vinylphenol was shown to inhibit metastasis and cancer stemness in CSC-enriched breast cancer cells. Since conventional therapies not targeting CSCs possibly lead to failure to eliminate cancer, 4-vinylphenol is a highly potential therapeutic agent for breast cancer patients.


Cancer stem-like cells Breast cancer 4-Vinylphenol Metastasis Drug resistance Tumorigenicity 





Aldehyde dehydrogenase


Cancer stem-like cell


Chemokine receptor-type 4


Extracellular matrix


Epidermal growth factor receptor


Epithelial–mesenchymal transition


Focal adhesion kinase


Matrix metalloproteinase


3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide


Tissue inhibitor of matrix metalloproteinase


Phosphatidylinositol 3-kinase


Urokinase-type plasminogen activator



The authors would like to thank Mr. Ching-Po Lau, Miss Xiaoxiao Wu and Miss Julia Lee for their technical support.

Author contributions

HWL designed the study, performed in vitro and in vivo experiments, analyzed data, and drafted the manuscript; CHK and GGLY designed the study and revised the manuscript; IH provided technical support on cancer stem cell study and CBSL was in-charge and supervised the project and revised the manuscript. All authors discussed the results and commented on the manuscript at all stages.


This study was partly supported by grants of the State Key Laboratory of Phytochemistry and Plant Resources in West China (CUHK) from Innovation and Technology Commission, HKSAR, and the Chinese University of Hong Kong.

Compliance with ethical standards

Conflict of interest

Dr. Leung declares that she has no conflict of interest. Dr. Ko declares that he has no conflict of interest. Dr. Yue declares that she has no conflict of interest. Prof. Herr declares that she has no conflict of interest. Prof. Lau declares that she has no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All experimental protocols were approved by the Animal Experimentation Ethics Committee of The Chinese University of Hong Kong with reference numbers Ref No. 15/128/MIS and 16/162/MIS.

Research involving human and animal participants

This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

280_2018_3601_MOESM1_ESM.docx (177 kb)
Supplementary material 1 (DOCX 176 KB)


  1. 1.
    Visvader JE, Lindeman GJ (2012) Cancer stem cells: current status and evolving complexities. Cell Stem Cell 10:717–728CrossRefPubMedGoogle Scholar
  2. 2.
    Lee CJ, Dosch J, Simeone DM (2008) Pancreatic cancer stem cells. J Clin Oncol 26(17):2806–2812CrossRefPubMedGoogle Scholar
  3. 3.
    Tanei T, Morimoto K, Shimazu K, Kim SJ, Tanji Y, Tahuchi T et al (2009) Association of breast cancer stem cells identified by aldehyde dehydrogenase 1 expression with resistance to sequential Paclitaxel and Epirubicin-based chemotherapy for breast cancers. Clin Cancer Res 15:4234–4241CrossRefPubMedGoogle Scholar
  4. 4.
    Siegel R, Ma J, Zou Z, Jemal A (2014) Cancer statistics 2014. CA Cancer J Clin 64:9–29CrossRefPubMedGoogle Scholar
  5. 5.
    Brewster AM, Hortobagyi GN, Broglio KR, Kau SW, Santa-Maria CA, Buzdar AU et al (2008) Residual risk of breast cancer recurrence 5 years after adjuvant therapy. J Natl Cancer Inst 100:1179–1183CrossRefPubMedGoogle Scholar
  6. 6.
    Mimeault M, Batra SK (2010) New advances on critical implications of tumour- and metastasis-initiating cells in cancer progression, treatment resistance and disease recurrence. Histol Histopathol 25(8):1057–1073PubMedPubMedCentralGoogle Scholar
  7. 7.
    Steeg PS (2006) Tumor metastasis: mechanistic insights and clinical challenges. Nat Med 12(8):895–904CrossRefPubMedGoogle Scholar
  8. 8.
    Muller A, Homey B, Soto H, Ge N, Catron D, Buchanan ME et al (2001) Involvement of chemokine receptors in breast cancer metastasis. Nature 410:50–56CrossRefPubMedGoogle Scholar
  9. 9.
    Chen R, He J, Tong X, Tang L, Liu M (2016) The Hedyotis diffusa Willd. (Rubiaceae): a review on phytochemistry, pharmacology, quality control and pharmacokinetics. Molecules 21(6):E710CrossRefPubMedGoogle Scholar
  10. 10.
    Withycombe DA, Lindsay RC, Stuiber DA (1978) Isolation and identification of volatile components from wild rice grain (Zizania aquatica). J Agric Food Chem 26:816–822CrossRefGoogle Scholar
  11. 11.
    Fras P, Campos FM, Hogg T, Couto JA (2014) Production of volatile phenols by Lactobacillus plantarum in wine conditions. Biotechnol Lett 36:281–285CrossRefPubMedGoogle Scholar
  12. 12.
    Godoy L, Garrido D, Martínez C, Saavedra J, Combina M, Ganga MA (2009) Study of the coumarate decarboxylase and vinylphenol reductase activities of Dekkera bruxellensis (anamorph Brettanomyces bruxellensis) isolates. Lett Appl Microbiol 48(4):452–457CrossRefPubMedGoogle Scholar
  13. 13.
    Chen XZ, Cao ZY, Chen TS, Zhang YQ, Liu ZZ, Su YT et al (2012) Water extract of Hedyotis diffusa Willd suppresses proliferation of human HepG2 cells and potentiates the anticancer efficacy of low-dose 5-fluorouracil by inhibiting the CDK2-E2F1 pathway. Oncol Rep 28:742–748CrossRefPubMedGoogle Scholar
  14. 14.
    Lin J, Wei L, Shen A, Cai Q, Xu W, Li H et al (2013) Hedyotis diffusa Willd extract suppress Sonic hedgehog signaling leading to the inhibition of colorectal cancer angiogenesis. Int J Oncol 42(2):651–656CrossRefPubMedGoogle Scholar
  15. 15.
    Li R, Zhao H, Lin Y (2002) Anti-tumour effect and protective effect on chemotherapeutic damage of water soluble extracts from Hedyotis diffusa. J Chin Pharmaceut Sci 11(2):54–58Google Scholar
  16. 16.
    Yue GGL, Lee JKM, Kwok HF, Cheng L, Wong ECW, Jiang L et al (2015) Novel PI3K/AKT targeting anti-angiogenic activities of 4-vinylphenol, a new therapeutic potential of a well-known styrene metabolite. Sci Rep 5:11149CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Leung HW, Wang Z, Yue GGL, Zhao SM, Lee JK, Fung KP et al (2015) Cyclopeptide RA-V inhibits cell adhesion and invasion in both estrogen receptor positive and negative breast cancer cells via PI3K/AKT and NF-κB signaling pathways. Biochim Biophys Acta 1853(8):1827–1840CrossRefPubMedGoogle Scholar
  18. 18.
    Kalluri R, Weinberg RA (2009) The basics of epithelial-mesenchymal transition. J Clin Investig 119(6):1420–1428CrossRefPubMedGoogle Scholar
  19. 19.
    Al-Hajj M, Wicha MS, Benito-Hernanderz A, Morrison SJ, Clarke MF (2003) Prospective identification of tumourigenic breast cancer cells. Proc Natl Acad Sci USA 100:3983–3988Google Scholar
  20. 20.
    Charafe-Jauffret E, Ginestier C, Lovino F, Tarpin C, Diebel M, Esterni B et al Birnbaum D, Viens P, Wicha MS (2010) Aldehyde dehydrogenase 1-positive cancer stem cells mediate metastasis and poor clinical outcome in inflammatory breast cancer. Clin Cancer Res 16(1):45–55CrossRefPubMedGoogle Scholar
  21. 21.
    Croker AK, Goodale D, Chu J, Postenka C, Hedley BD, Hess DA et al (2009) High aldehyde dehydrogenase and expression of cancer stem cell markers selects for breast cancer cells with enhanced malignant and metastatic ability. J Cell Mol Med 13(8B):2236–2252CrossRefPubMedGoogle Scholar
  22. 22.
    Marcato P, Dean CA, Pan D, Araslanova R, Cillis M, Joshi M et al (2011) Aldehyde dehydrogenase activity of breast cancer stem cells is primarily due to isoform ALDH1A3 and its expression is predictive of metastasis. Stem Cells 29(1):32–45CrossRefPubMedGoogle Scholar
  23. 23.
    Gupta PB, Onder TT, Jiang G, Tao K, Kuperwasser C, Weinberg RA et al (2009) Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell 138(4):645–659CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Mare JA, Sterrenberg JN, Sukhthankar MG, Chiwakata MT, Beukes DR, Blatch GL et al (2013) Assessment of potential anti-cancer stem cell activity of marine algal compounds using in vitro mammosphere assay. Cancer Cell Int 13(1):39CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Hermann PC, Huber SL, Herrler T, Aicher A, Ellwart JW, Guba M et al (2007) Distinct populations of cancer stem cells determine tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 1:313–323CrossRefPubMedGoogle Scholar
  26. 26.
    Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY et al (2008) The epithelial-mesenchymal transition generate cells with properties of stem cells. Cell 133(4):704–715CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Sheridan C, Kishimoto H, Fuchs RK, Mehrotra S, Bhat-Nakshatri P, Turner CH et al (2006) CD44+/CD24 breast cancer cells exhibit enhanced invasive properties: an early step necessary for metastasis. Breast Cancer Res 8(5):R59CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Yang J, Weinberg RA (2008) Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis. Dev Cell 14(6):818–829CrossRefPubMedGoogle Scholar
  29. 29.
    Teng Y, Xie X, Walker S, White DT, Mumm JS, Cowell JK (2013) Evaluating human cancer cell metastasis in zebrafish. BMC Cancer 13:453CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of Chinese MedicineThe Chinese University of Hong KongShatinHong Kong
  2. 2.State Key Laboratory of Phytochemistry and Plant Resources in West ChinaThe Chinese University of Hong KongShatinHong Kong
  3. 3.Department of General, Visceral and Transplantation SurgeryUniversity of HeidelbergHeidelbergGermany

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