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Horm Metab Res 2014; 46(11): 753-760
DOI: 10.1055/s-0034-1376977
Endocrine Research
© Georg Thieme Verlag KG Stuttgart · New York

Formononetin Inhibits Migration and Invasion of MDA-MB-231 and 4T1 Breast Cancer Cells by Suppressing MMP-2 and MMP-9 Through PI3K/AKT Signaling Pathways

R. Zhou   *
1   Department of Chest and Breast Surgery, Xiamen Hospital of Traditional Chinese Medicine, Fujian University of Traditional Chinese Medicine, Xiamen, P. R. China
,
L. Xu   *
2   Department of Orthopedic Oncology, Changzheng Hospital, Second Military Medical University, Shanghai, P. R. China
,
M. Ye
3   Department of Breast Surgery, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, P. R. China
,
M. Liao
3   Department of Breast Surgery, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, P. R. China
,
H. Du
1   Department of Chest and Breast Surgery, Xiamen Hospital of Traditional Chinese Medicine, Fujian University of Traditional Chinese Medicine, Xiamen, P. R. China
,
H. Chen
3   Department of Breast Surgery, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, P. R. China
› Author Affiliations
Further Information

Publication History

received 19 September 2013

accepted 05 May 2014

Publication Date:
30 June 2014 (online)

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Abstract

Formononetin is a naturally existing isoflavone, which can be found in the roots of Astragalus membranaceus, Trifolium pratense, Glycyrrhiza glabra, and Pueraria lobata. It was found to be associated with inhibition of cell proliferation and cell cycle progression, as well as induction of apoptosis in various cancer cell lines. However, the effect of formononetin on breast cancer cell metastasis remains unclear. In this study, we examined the effect of formononetin on the migration and invasion of breast cancer cells MDA-MB-231 and 4T1 in vitro and in vivo. Our data demonstrated that formononetin did not effectively inhibit the cell viability of MDA-MB-231 and 4T1 in 24 h with the concentration lower than 160 μmol/l. When treated with nontoxic concentration of formononetin, the migration and invasion of MDA-MB-231 and 4T1 cells were markedly suppressed by wound healing assay, chamber invasion assay, and in vivo mouse metastasis model. In vitro, formononetin reduced the expression of matrix metalloproteinase-2 (MMP-2), MMP-9 and increased the expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2. Furthermore, the immunofluorescence and immunoblotting assays indicated that formononetin was very effective in suppressing the phosphorylation of Akt and PI3K. Collectively, these results suggest that formononetin inhibited breast cancer cell migration and invasion by reducing the expression of MMP-2 and MMP-9 through the PI3K/AKT signaling pathway. These findings demonstrate a potentially new therapeutic strategy of formononetin as anti-invasive agent for breast cancer.

Key words

formononetin - breast cancer - migration - PI3K/AKT - matrix metalloproteinases

* R. Zhou and L. Xu contributed equally to this study and should be considered as first co-authors.


 

  • References

  • 1 Redig AJ, McAllister SS. Breast cancer as a systemic disease: a view of metastasis. J Intern Med 2013; 274: 113-126
    CrossrefPubMedGoogle Scholar
  • 2 Mahamodhossen YA, Liu W, Rong-Rong Z. Triple-negative breast cancer: new perspectives for novel therapies. Med Oncol 2013; 30: 653
    CrossrefPubMedGoogle Scholar
  • 3 Amos KD, Adamo B, Anders CK. Triple-negative breast cancer: an update on neoadjuvant clinical trials. Int J Breast Cancer 2012; 2012: 385978
    PubMedGoogle Scholar
  • 4 Baser O, Wei W, Xie L, Henk HJ, Teitelbaum A. Retrospective study of patients with metastatic triple-negative breast cancer: survival, health care utilization, and cost. Comm Oncol 2012; 9: 8-14
    PubMedGoogle Scholar
  • 5 Talvensaari-Mattila A, Paakko P, Hoyhtya M, Blanco-Sequeiros G, Turpeenniemi-Hujanen T. Matrix metalloproteinase-2 immunoreactive protein: a marker of aggressiveness in breast carcinoma. Cancer 1998; 83: 1153-1162
    CrossrefPubMedGoogle Scholar
  • 6 Pellikainen JM, Ropponen KM, Kataja VV, Kellokoski JK, Eskelinen MJ, Kosma VM. Expression of matrix metalloproteinase (MMP)-2 and MMP-9 in breast cancer with a special reference to activator protein-2, HER2, and prognosis. Clinical cancer research 2004; 10: 7621-7628
    CrossrefPubMedGoogle Scholar
  • 7 Li HC, Cao DC, Liu Y, Hou YF, Wu J, Lu JS, Di GH, Liu G, Li FM, Ou ZL, Jie C, Shen ZZ, Shao ZM. Prognostic value of matrix metalloproteinases (MMP-2 and MMP-9) in patients with lymph node-negative breast carcinoma. Breast Cancer Res Treat 2004; 88: 75-85
    CrossrefPubMedGoogle Scholar
  • 8 Egeblad M, Werb Z. New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2002; 2: 161-174
    CrossrefPubMedGoogle Scholar
  • 9 Li W, Sun YN, Yan XT, Yang SY, Kim S, Lee YM, Koh YS, Kim YH. Flavonoids from Astragalus membranaceus and their inhibitory effects on LPS-stimulated pro-inflammatory cytokine production in bone marrow-derived dendritic cells. Arch Pharm Res 2014; 37: 186-192
    CrossrefPubMedGoogle Scholar
  • 10 Ma Z, Ji W, Fu Q, Ma S. Formononetin Inhibited the Inflammation of LPS-Induced Acute Lung Injury in Mice Associated with Induction of PPAR Gamma Expression. Inflammation 2013; 36: 1560-1566
    CrossrefPubMedGoogle Scholar
  • 11 Lai PK, Chan JY, Cheng L, Lau CP, Han SQ, Leung PC, Fung KP, Lau CB. Isolation of anti-inflammatory fractions and compounds from the root of Astragalus membranaceus. Phytother Res 2013; 27: 581-587
    CrossrefPubMedGoogle Scholar
  • 12 Ren J, Xu HJ, Cheng H, Xin WQ, Chen X, Hu K. Synthesis and antitumor activity of Formononetin nitrogen mustard derivatives. Eur J Med Chem 2012; 54: 175-187
    CrossrefPubMedGoogle Scholar
  • 13 Ji ZN, Zhao WY, Liao GR, Choi RC, Lo CK, Dong TT, Tsim KW. In vitro estrogenic activity of Formononetin by two bioassay systems. Gynecological endocrinology: the official journal of the International Society of Gynecol Endocrinol 2006; 22: 578-584
    PubMedGoogle Scholar
  • 14 Park J, Kim SH, Cho D, Kim TS. Formononetin, a phyto-oestrogen, and its metabolites up-regulate interleukin-4 production in activated T cells via increased AP-1 DNA binding activity. Immunology 2005; 116: 71-81
    CrossrefPubMedGoogle Scholar
  • 15 Sun T, Wang J, Huang LH, Cao YX. Antihypertensive effect of Formononetin through regulating the expressions of eNOS, 5-HT2A/1B receptors and alpha1-adrenoceptors in spontaneously rat arteries. Eur J Pharmacol 2013; 699: 241-249
    CrossrefPubMedGoogle Scholar
  • 16 Ye Y, Hou R, Chen J, Mo L, Zhang J, Huang Y, Mo Z. Formononetin-induced apoptosis of human prostate cancer cells through ERK1/2 mitogen-activated protein kinase inactivation. Horm Metab Res 2012; 44: 263-267
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Chen J, Zeng J, Xin M, Huang W, Chen X. Formononetin induces cell cycle arrest of human breast cancer cells via IGF1/PI3K/Akt pathways in vitro and in vivo. Horm Metab Res 2011; 43: 681-686
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Chen J, Sun L. Formononetin-induced apoptosis by activation of Ras/p38 mitogen-activated protein kinase in estrogen receptor-positive human breast cancer cells. Horm Metab Res 2012; 44: 943-948
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Mansoor TA, Ramalho RM, Luo X, Ramalhete C, Rodrigues CM, Ferreira MJ. Isoflavones as apoptosis inducers in human hepatoma HuH-7 cells. Phytother Res 2011; 25: 1819-1824
    CrossrefPubMedGoogle Scholar
  • 20 Auyeung KK, Law PC, Ko JK. Novel anti-angiogenic effects of Formononetin in human colon cancer cells and tumor xenograft. Oncol Rep 2012; 28: 2188-2194
    PubMedGoogle Scholar
  • 21 Krenn L, Paper DH. Inhibition of angiogenesis and inflammation by an extract of red clover (Trifolium pratense L.). Phytomedicine 2009; 16: 1083-1088
    CrossrefPubMedGoogle Scholar
  • 22 Chavez KJ, Garimella SV, Lipkowitz S. Triple negative breast cancer cell lines: one tool in the search for better treatment of triple negative breast cancer. Breast Dis 2010; 32: 35-48
    PubMedGoogle Scholar
  • 23 Heppner GH, Miller FR, Shekhar PM. Nontransgenic models of breast cancer. Breast Cancer Res 2000; 2: 331-334
    CrossrefPubMedGoogle Scholar
  • 24 Tao K, Fang M, Alroy J, Sahagian GG. Imagable 4T1 model for the study of late stage breast cancer. BMC Cancer 2008; 8: 228
    CrossrefPubMedGoogle Scholar
  • 25 Tyagi AM, Srivastava K, Singh AK, Kumar A, Changkija B, Pandey R, Lahiri S, Nagar GK, Yadav DK, Maurya R, Trivedi R, Singh D. Formononetin reverses established osteopenia in adult ovariectomized rats. Menopause (New York, NY) 2012; 19: 856-863
    CrossrefPubMedGoogle Scholar
  • 26 Scully OJ, Bay BH, Yip G, Yu Y. Breast cancer metastasis. Cancer Genom Proteom 2012; 9: 311-320
    PubMedGoogle Scholar
  • 27 Alix-Panabieres C, Muller V, Pantel K. Current status in human breast cancer micrometastasis. Curr Opin Oncol 2007; 19: 558-563
    CrossrefPubMedGoogle Scholar
  • 28 Deryugina EI, Quigley JP. Matrix metalloproteinases and tumor metastasis. Cancer Metas Rev 2006; 25: 9-34
    PubMedGoogle Scholar
  • 29 Jezierska A, Motyl T. Matrix metalloproteinase-2 involvement in breast cancer progression: a mini-review. Med Sci Mon 2009; 15: RA32-RA40
    PubMedGoogle Scholar
  • 30 Vizoso FJ, Gonzalez LO, Corte MD, Rodriguez JC, Vazquez J, Lamelas ML, Junquera S, Merino AM, Garcia-Muniz JL. Study of matrix metalloproteinases and their inhibitors in breast cancer. Brit J Cancer 2007; 96: 903-911
    CrossrefPubMedGoogle Scholar
  • 31 Hua J, Muschel RJ. Inhibition of matrix metalloproteinase 9 expression by a ribozyme blocks metastasis in a rat sarcoma model system. Cancer research 1996; 56: 5279-5284
    PubMedGoogle Scholar
  • 32 Kondraganti S, Mohanam S, Chintala SK, Kin Y, Jasti SL, Nirmala C, Lakka SS, Adachi Y, Kyritsis AP, Ali-Osman F, Sawaya R, Fuller GN, Rao JS. Selective suppression of matrix metalloproteinase-9 in human glioblastoma cells by antisense gene transfer impairs glioblastoma cell invasion. Cancer research 2000; 60: 6851-6855
    PubMedGoogle Scholar
  • 33 Iurlaro M, Loverro G, Vacca A, Cormio G, Ribatti D, Minischetti M, Ria R, Bruno M, Selvaggi L. Angiogenesis extent and expression of matrix metalloproteinase-2 and -9 correlate with upgrading and myometrial invasion in endometrial carcinoma. Eur J Clin Invest 1999; 29: 793-801
    CrossrefPubMedGoogle Scholar
  • 34 Cruz-Munoz W, Khokha R. The role of tissue inhibitors of metalloproteinases in tumorigenesis and metastasis. Crit Rev Clin Lab Sci 2008; 45: 291-338
    CrossrefPubMedGoogle Scholar
  • 35 Khokha R. Suppression of the tumorigenic and metastatic abilities of murine B16-F10 melanoma cells in vivo by the overexpression of the tissue inhibitor of the metalloproteinases-1. J Nat Cancer Inst 1994; 86: 299-304
    CrossrefPubMedGoogle Scholar
  • 36 Khokha R, Zimmer MJ, Graham CH, Lala PK, Waterhouse P. Suppression of invasion by inducible expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) in B16-F10 melanoma cells. J Nat Cancer Inst 1992; 84: 1017-1022
    CrossrefPubMedGoogle Scholar
  • 37 Valente P, Fassina G, Melchiori A, Masiello L, Cilli M, Vacca A, Onisto M, Santi L, Stetler-Stevenson WG, Albini A. TIMP-2 over-expression reduces invasion and angiogenesis and protects B16F10 melanoma cells from apoptosis. Int J Cancer 1998; 75: 246-253
    CrossrefPubMedGoogle Scholar
  • 38 Chen JS, Wang Q, Fu XH, Huang XH, Chen XL, Cao LQ, Chen LZ, Tan HX, Li W, Bi J, Zhang LJ. Involvement of PI3K/PTEN/AKT/mTOR pathway in invasion and metastasis in hepatocellular carcinoma: Association with MMP-9. Hepatol Res 2009; 39: 177-186
    CrossrefPubMedGoogle Scholar
  • 39 Li X, Yang Z, Song W, Zhou L, Li Q, Tao K, Zhou J, Wang X, Zheng Z, You N, Dou K, Li H. Overexpression of Bmi-1 contributes to the invasion and metastasis of hepatocellular carcinoma by increasing the expression of matrix metalloproteinase (MMP)2, MMP-9 and vascular endothelial growth factor via the PTEN/PI3K/Akt pathway. Int J Oncol 2013; 43: 793-802
    PubMedGoogle Scholar
  • 40 Tian T, Nan KJ, Guo H, Wang WJ, Ruan ZP, Wang SH, Liang X, Lu CX. PTEN inhibits the migration and invasion of HepG2 cells by coordinately decreasing MMP expression via the PI3K/Akt pathway. Oncol Rep 2010; 23: 1593-1600
    PubMedGoogle Scholar
  • 41 Qiu Q, Yang M, Tsang BK, Gruslin A. EGF-induced trophoblast secretion of MMP-9 and TIMP-1 involves activation of both PI3K and MAPK signalling pathways. Reproduction 2004; 128: 355-363
    CrossrefPubMedGoogle Scholar
  • 42 Ye B, Jiang LL, Xu HT, Zhou DW, Li ZS. Expression of PI3K/AKT pathway in gastric cancer and its blockade suppresses tumor growth and metastasis. Int J Immunopathol Pharmacol 2012; 25: 627-636
    PubMedGoogle Scholar
  • 43 Huang CY, Fong YC, Lee CY, Chen MY, Tsai HC, Hsu HC, Tang CH. CCL5 increases lung cancer migration via PI3K, Akt and NF-kappaB pathways. Biochem Pharmacol 2009; 77: 794-803
    CrossrefPubMedGoogle Scholar
  • 44 Zhang ZJ, Ma SL. miRNAs in breast cancer tumorigenesis (Review). Oncol Rep 2012; 27: 903-910
    PubMedGoogle Scholar
  • 45 Jung CH, Kim EM, Park JK, Hwang SG, Moon SK, Kim WJ, Um HD. Bmal1 suppresses cancer cell invasion by blocking the phosphoinositide 3-kinase-Akt-MMP-2 signaling pathway. Oncol Rep 2013; 29: 2109-2113
    PubMedGoogle Scholar
  • 46 Xie Y, Abel PW, Kirui JK, Deng C, Sharma P, Wolff DW, Toews ML, Tu Y. Identification of upregulated phosphoinositide 3-kinase gamma as a target to suppress breast cancer cell migration and invasion. Biochem Pharmacol 2013; 85: 1454-1462
    CrossrefPubMedGoogle Scholar
  • 47 Karam AK, Santiskulvong C, Fekete M, Zabih S, Eng C, Dorigo O. Cisplatin and PI3kinase inhibition decrease invasion and migration of human ovarian carcinoma cells and regulate matrix-metalloproteinase expression. Cytoskeleton 2010; 67: 535-544
    PubMedGoogle Scholar
  • 48 Chen J, Lan T, Hou J, Zhang J, An Y, Tie L, Pan Y, Liu J, Li X. Atorvastatin sensitizes human non-small cell lung carcinomas to carboplatin via suppression of AKT activation and upregulation of TIMP-1. Int J Biochem Cell Biol 2012; 44: 759-769
    CrossrefPubMedGoogle Scholar
  • 49 Ko HS, Lee HJ, Kim SH, Lee EO. Piceatannol suppresses breast cancer cell invasion through the inhibition of MMP-9: involvement of PI3K/AKT and NF-kappaB pathways. J Agric Food Chem 2012; 60: 4083-4089
    CrossrefPubMedGoogle Scholar
  • 50 Lee H, Kim JS, Kim E. Fucoidan from seaweed Fucus vesiculosus inhibits migration and invasion of human lung cancer cell via PI3K-Akt-mTOR pathways. PloS One 2012; 7: e50624
    CrossrefPubMedGoogle Scholar
  • 51 Hsieh CY, Tsai PC, Chu CL, Chang FR, Chang LS, Wu YC, Lin SR. Brazilein suppresses migration and invasion of MDA-MB-231 breast cancer cells. Chemicobiol Interact 2013; 204: 105-115
    PubMedGoogle Scholar
  • 52 Tang L, Ma X, Tian Q, Cheng Y, Yao H, Liu Z, Qu X, Han X. Inhibition of angiogenesis and invasion by DMBT is mediated by downregulation of VEGF and MMP-9 through Akt pathway in MDA-MB-231 breast cancer cells. Food Chem Toxicol 2013; 56: 204-213
    CrossrefPubMedGoogle Scholar
  • 53 Tong B, Lu D, Wei Z, Wang T, Xia Y, Dai Y. Gleditsioside B, a triterpene saponin isolated from the anomalous fruits of Gleditsia sinensis Lam., abrogates bFGF-induced endothelial cell migration through preventing the activation of MMP-2 and FAK via inhibiting ERK and PI3K/AKT signaling pathways. Vascul Pharmacol 2013; 58: 118-126
    CrossrefPubMedGoogle Scholar
  • 54 Yan KH, Lee LM, Yan SH, Huang HC, Li CC, Lin HT, Chen PS. Tomatidine inhibits invasion of human lung adenocarcinoma cell A549 by reducing matrix metalloproteinases expression. Chemico-biological interactions 2013; 203: 580-587
    PubMedGoogle Scholar
  • 55 Chen SM, Liu JL, Wang X, Liang C, Ding J, Meng LH. Inhibition of tumor cell growth, proliferation and migration by X-387, a novel active-site inhibitor of mTOR. Biochem Pharmacol 2012; 83: 1183-1194
    CrossrefPubMedGoogle Scholar
  • 56 Li M, Wu S, Liu Z, Zhang W, Xu J, Wang Y, Liu J, Zhang D, Tian H, Li Y, Ye W. Arenobufagin, a bufadienolide compound from toad venom, inhibits VEGF-mediated angiogenesis through suppression of VEGFR-2 signaling pathway. Biochem Pharmacol 2012; 83: 1251-1260
    CrossrefPubMedGoogle Scholar