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01-02-2015 | Preclinical Study

The role of polycomb group protein Bmi-1 and Notch4 in breast cancer stem cell inhibition by benzyl isothiocyanate

Authors: Su-Hyeong Kim, Shivendra V. Singh

Published in: Breast Cancer Research and Treatment | Issue 3/2015

Abstract

We showed previously that garden cress constituent benzyl isothiocyanate (BITC) inhibits self-renewal of breast cancer stem cells (bCSC) in vitro and in vivo. The present study offers novel insights into the mechanism by which BITC inhibits bCSC. Flow cytometry and mammosphere assay were performed to quantify bCSC fraction. Protein expression was determined by western blotting. Apoptosis was assessed by flow cytometry using Annexin V-propidium iodide method. Cell migration was determined by Boyden chamber assay. BITC treatment resulted in a marked decrease in protein level of polycomb group protein B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1) in cultured human breast cancer cells (MCF-7, SUM159, MDA-MB-231, and MDA-MB-361) and MDA-MB-231 xenografts in vivo. Overexpression (MCF-7) or knockdown (SUM159, and MDA-MB-231) of Bmi-1 protein had no meaningful impact on the BITC’s ability to inhibit cell viability and cell migration and/or induce apoptosis. On the other hand, inhibition of bCSC markers (aldehyde dehydrogenase 1 activity and mammosphere frequency) resulting from BITC exposure was significantly altered by Bmi-1 overexpression and knockdown. BITC was previously shown to cause activation of Notch1, Notch2, and Notch4 in association with induction of γ-secretase complex component Nicastrin, which are also implicated in maintenance of cancer stemness. BITC-mediated inhibition of bCSC was augmented by knockdown of Notch4 and Nicastrin, but not by RNA interference of Notch1 or Notch2. The present study highlights important roles for Bmi-1 and Notch4 in BITC-mediated suppression of bCSC.

Fisher B, Costantino JP, Wickerham DL, Redmond CK, Kavanah M, Cronin WM, Vogel V, Robidoux A, Dimitrov N, Atkins J, Daly M, Wieand S, Tan-Chiu E, Ford L, Wolmark N (1998) Tamoxifen for prevention of breast cancer: report of the National Surgical Adjuvant Breast and Bowel Project P-1 study. J Natl Cancer Inst 90(18):1371–1388CrossRefPubMed

Goss PE, Ingle JN, Ales-Martinez JE, Cheung AM, Chlebowski RT, Wactawski-Wende J, McTierman A, Robbins J, Johnson KC, Martin LW, Winquist E, Sarto GE, Garber JE, Fabian CJ, Pujol P, Maunsell E, Farmer P, Gelmon KA, Tu D, Richardson H, NCIC CTG MAP.3 Study Investigators (2011) Exemestane for breast-cancer prevention in postmenopausal women. N Eng J Med 364(25):2381–2391CrossRef

Higgins MJ, Baselga J (2011) Breast cancer in 2010: novel targets and therapies for a personalized approach. Nat Rev Clin Oncol 8(2):65–66CrossRefPubMed

van Zitteren M, van der Net JB, Kundu S, Freedman AN, van Duijn CM, Janssens AC (2011) Genome-based prediction of breast cancer risk in the general population: a modeling study based on meta-analyses of genetic associations. Cancer Epidemiol Biomarkers Prev 20(1):9–22CrossRefPubMed

Siegel R, Naishadham D, Jemal A (2013) Cancer statistics, 2013. CA Cancer J Clin 63(1):11–30CrossRefPubMed

Velasco-Velázquez MA, Homsi N, De La Fuente M, Pestell RG (2012) Breast cancer stem cells. Int J Biochem Cell Biol 44(4):573–577CrossRefPubMedCentralPubMed

O’Brien CS, Farnie G, Howell SJ, Clarke RB (2011) Breast cancer stem cells and their role in resistance to endocrine therapy. Horm Cancer 2(2):91–103CrossRefPubMed

Surh YJ (2003) Cancer chemoprevention with dietary phytochemicals. Nat Rev Cancer 3(10):768–780CrossRefPubMed

Vyas AR, Singh SV (2014) Molecular targets and mechanisms of cancer prevention and treatment by withaferin A, a naturally occurring steroidal lactone. AAPS J 16(1):1–10CrossRefPubMedCentralPubMed

10.

Sehrawat A, Singh SV (2013) Molecular mechanisms of cancer chemoprevention with benzyl isothiocyanate. In: Kong AN (ed) Inflammation, oxidative stress, and cancer. Taylor & Francis, New York, pp 447–462. doi:10.1201/b15323-32 CrossRef

11.

Ambrosone CB, McCann SE, Freudenheim JL, Marshall JR, Zhang Y, Shields PG (2004) Breast cancer risk in premenopausal women is inversely associated with consumption of broccoli, a source of isothiocyanates, but is not modified by GST genotype. J Nutr 134(5):1134–1138PubMed

12.

Wattenberg LW (1977) Inhibition of carcinogenic effects of polycyclic hydrocarbons by benzyl isothiocyanate and related compounds. J Natl Cancer Inst 58(2):395–398PubMed

13.

Warin R, Xiao D, Arlotti JA, Bommareddy A, Singh SV (2010) Inhibition of human breast cancer xenograft growth by cruciferous vegetable constituent benzyl isothiocyanate. Mol Carcinog 49(5):500–507CrossRefPubMedCentralPubMed

14.

Kim EJ, Hong JE, Eom SJ, Lee JY, Park JH (2011) Oral administration of benzyl-isothiocyanate inhibits solid tumor growth and lung metastasis of 4T1 murine mammary carcinoma cells in BALB/c mice. Breast Cancer Res Treat 130(1):61–71CrossRefPubMed

15.

Warin R, Chambers WH, Potter DM, Singh SV (2009) Prevention of mammary carcinogenesis in MMTV-neu mice by cruciferous vegetable constituent benzyl isothiocyanate. Cancer Res 69(24):9473–9480CrossRefPubMedCentralPubMed

16.

Kim SH, Sehrawat A, Singh SV (2013) Dietary chemopreventative benzyl isothiocyanate inhibits breast cancer stem cells in vitro and in vivo. Cancer Prev Res 6(8):782–790CrossRef

17.

Liu S, Dontu G, Mantle ID, Patel S, Ahn NS, Jackson KW, Suri P, Wicha MS (2006) Hedgehog signaling and Bmi-1 regulate self-renewal of normal and malignant human mammary stem cells. Cancer Res 66(12):6063–6071CrossRefPubMed

18.

Harrison H, Farnie G, Howell SJ, Rock RE, Stylianou S, Brennan KR, Bundred NJ, Clarke RB (2010) Regulation of breast cancer stem cell activity by signaling through the Notch4 receptor. Cancer Res 70(2):709–718CrossRefPubMedCentralPubMed

19.

Paranjape AN, Balaji SA, Mandal T, Krushik EV, Nagaraj P, Mukherjee G, Rangarajan A (2014) Bmi1 regulates self-renewal and epithelial to mesenchymal transition in breast cancer cells through Nanog. BMC Cancer 14:785CrossRefPubMedCentralPubMed

20.

Gonzalez ME, Moore HM, Li X, Toy KA, Huang W, Sabel MS, Kidwell KM, Kleer CG (2014) EZH2 expands breast stem cells through activation of NOTCH1 signaling. Proc Natl Acad Sci USA 111(8):3098–3103CrossRefPubMedCentralPubMed

21.

Kim SH, Sehrawat A, Singh SV (2012) Notch2 activation by benzyl isothiocyanate impedes its inhibitory effect on breast cancer cell migration. Breast Cancer Res Treat 134(3):1067–1079CrossRefPubMedCentralPubMed

22.

Xiao D, Srivastava SK, Lew KL, Zeng Y, Hershberger P, Johnson CS, Trump DL, Singh SV (2003) Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits proliferation of human prostate cancer cells by causing G₂/M arrest and inducing apoptosis. Carcinogenesis 24(5):891–897CrossRefPubMed

23.

Xiao D, Lew KL, Kim YA, Zeng Y, Hahm ER, Dhir R, Singh SV (2006) Diallyl trisulfide suppresses growth of PC-3 human prostate cancer xenograft in vivo in association with Bax and Bak induction. Clin Cancer Res 12(22):6836–6843CrossRefPubMed

24.

Sakao K, Singh SV (2012) d,l-Sulforaphane-induced apoptosis in human breast cancer cells is regulated by the adapter protein p66^Shc. J Cell Biochem 113(2):599–610CrossRefPubMedCentralPubMed

25.

Finn RS, Dering J, Conklin D, Kalous O, Cohen DJ, Desai AJ, Ginther C, Atefi M, Chen I, Fowst C, Los G, Slamon DJ (2009) PD0332991, a selective cyclin D1 kinase 4/6 inhibitor, preferentially inhibits proliferation and luminal estrogen receptor-positive human breast cancer cell lines in vitro. Breast Cancer Res 11(5):R77CrossRefPubMedCentralPubMed

26.

Boreddy SR, Pramanik KC, Srivastava SK (2011) Pancreatic tumor suppression by benzyl isothiocyanate is associated with inhibition of P13K/AKT/FOXO pathway. Clin Cancer Res 17(7):1784–1795CrossRefPubMedCentralPubMed

27.

Xu Z, Liu H, Lv X, Liu Y, Li S, Li H (2011) Knockdown of the Bmi-1 oncogene inhibits cell proliferation and induces cell apoptosis and is involved in the decrease of Akt phosphorylation in the human breast carcinoma cell line MCF-7. Oncol Rep 25(2):409–418PubMed

28.

Xiao D, Vogel V, Singh SV (2006) Benzyl isothiocyanate-induced apoptosis in human breast cancer cells is initiated by reactive oxygen species and regulated by Bax and Bak. Mol Cancer Ther 5(11):2931–2945CrossRefPubMed

29.

Wu XM, Liu X, Bu YQ, Sengupta J, Cui HJ, Yi FP, Liu T, Yuan CF, Shi YY, Song FZ (2009) RNAi-mediated silencing of the Bmi-1 gene causes growth inhibition and enhances doxorubicin-induced apoptosis in MCF-7 cells. Genet Mol Biol 32(4):697–703CrossRefPubMedCentralPubMed

30.

Guo BH, Feng Y, Zhang R, Xu LH, Li MZ, Kung HF, Song LB, Zeng MS (2011) Bmi-1 promotes invasion and metastasis, and its elevated expression is correlated with an advanced stage of breast cancer. Mol Cancer 10(1):10CrossRefPubMedCentralPubMed

31.

O’Neill CF, Urs S, Cinelli C, Lincoln A, Nadeau RJ, León R, Toher J, Mouta-Bellum C, Friesel RE, Liaw L (2007) Notch2 signaling induces apoptosis and inhibits human MDA-MB-231 xenograft growth. Am J Pathol 171(3):1023–1036CrossRefPubMedCentralPubMed

32.

Lombardo Y, Filipović A, Molyneux G, Periyasamy M, Giamas G, Hu Y, Trivedi PS, Wang J, Yagüe E, Michel L, Coombes RC (2012) Nicastrin regulates breast cancer stem cell properties and tumor growth in vitro and in vivo. Proc Natl Acad Sci USA 109(41):16558–16563CrossRefPubMedCentralPubMed

33.

Dimri GP, Martinez JL, Jacobs JJ, Keblusek P, Itahana K, Van Lohuizen M, Campisi J, Wazer DE, Band V (2002) The Bmi-1 oncogene induces telomerase activity and immortalizes human mammary epithelial cells. Cancer Res 62(16):4736–4745PubMed

34.

Wang Y, Zhe H, Ding Z, Gao P, Zhang N, Li G (2012) Cancer stem cell marker Bmi-1 expression is associated with basal-like phenotype and poor survival in breast cancer. World J Surg 36(5):1189–1194CrossRefPubMed

35.

Choi YJ, Choi YL, Cho EY, Shin YK, Sung KW, Hwang YK, Lee SJ, Kong G, Lee JE, Kim JS, Kim JH, Yang JH, Nam SJ (2009) Expression of Bmi-1 protein in tumor tissues is associated with favorable prognosis in breast cancer patients. Breast Cancer Res Treat 113(1):83–93CrossRefPubMed

36.

Rustighi A, Zannini A, Tiberi L, Sommaggio R, Piazza S, Sorrentino G, Nuzzo S, Tuscano A, Eterno V, Benvenuti F, Santarpia L, Aifantis I, Rosato A, Bicciato S, Zambelli A, Del Sal G (2014) Prolyl-isomerase Pin1 controls normal and cancer stem cells of the breast. EMBO Mol Med 6(1):99–119CrossRefPubMedCentralPubMed

Title: The role of polycomb group protein Bmi-1 and Notch4 in breast cancer stem cell inhibition by benzyl isothiocyanate
Authors: Su-Hyeong Kim
Shivendra V. Singh
Publication date: 01-02-2015
Publisher: Springer US
Published in: Breast Cancer Research and Treatment / Issue 3/2015
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-015-3279-5

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