Sulforaphane reduces molecular response to hypoxia in ovarian tumor cells independently of their resistance to chemotherapy

Pastorek,Michal; Simko,Veronika; Takacova,Martina; Barathova,Monika; Bartosova,Maria; Hunakova,Luba; Sedlakova,Olga; Hudecova,Sona; Krizanova,Olga; Dequiedt,Franck; Pastorekova,Silvia; Sedlak,Jan

doi:10.3892/ijo.2015.2987

Sulforaphane reduces molecular response to hypoxia in ovarian tumor cells independently of their resistance to chemotherapy

Authors:
- Michal Pastorek
- Veronika Simko
- Martina Takacova
- Monika Barathova
- Maria Bartosova
- Luba Hunakova
- Olga Sedlakova
- Sona Hudecova
- Olga Krizanova
- Franck Dequiedt
- Silvia Pastorekova
- Jan Sedlak
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Affiliations: Cancer Research Institute, Slovak Academy of Sciences, Bratislava, Slovak Republic, Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovak Republic, Institute of Molecular Physiology and Genetics, Slovak Academy of Sciences, Bratislava, Slovak Republic, Laboratory of Protein Signaling and Interactions, Interdisciplinary Cluster for Applied Genoproteomics, University of Liège, Sart-Tilman, Belgium
Published online on: May 5, 2015 https://doi.org/10.3892/ijo.2015.2987
Pages: 51-60
Copyright: © Pastorek et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

One of the recently emerging anticancer strategies is the use of natural dietary compounds, such as sulforaphane, a cancer-chemopreventive isothiocyanate found in broccoli. Based on the growing evidence, sulforaphane acts through molecular mechanisms that interfere with multiple oncogenic pathways in diverse tumor cell types. Herein, we investigated the anticancer effects of bioavailable concentrations of sulforaphane in ovarian carcinoma cell line A2780 and its two derivatives, adriamycin-resistant A2780/ADR and cisplatin-resistant A2780/CP cell lines. Since tumor microenvironment is characterized by reduced oxygenation that induces aggressive tumor phenotype (such as increased invasiveness and resistance to chemotherapy), we evaluated the effects of sulforaphane in ovarian cancer cells exposed to hypoxia (2% O2). Using the cell-based reporter assay, we identified several oncogenic pathways modulated by sulforaphane in hypoxia by activating anticancer responses (p53, ARE, IRF-1, Pax-6 and XRE) and suppressing responses supporting tumor progression (AP-1 and HIF-1). We further showed that sulforaphane decreases the level of HIF-1α protein without affecting its transcription and stability. It can also diminish transcription and protein level of the HIF-1 target, CA IX, which protects tumor cells from hypoxia-induced pH imbalance and facilitates their migration/invasion. Accordingly, sulforaphane treatment leads to diminished pH regulation and reduced migration of ovarian carcinoma cells. These effects occur in all three ovarian cell lines suggesting that sulforaphane can overcome the chemoresistance of cancer cells. This offers a path potentially exploitable in sensitizing resistant cancer cells to therapy, and opens a window for the combined treatments of sulforaphane either with conventional chemotherapy, natural compounds, or with other small molecules.

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1	Harris AL: Hypoxia - a key regulatory factor in tumor growth. Nat Rev Cancer. 2:38–47. 2002. View Article : Google Scholar : PubMed/NCBI
2	Semenza GL: Hypoxia-inducible factors: mediators of cancer progression and targets for cancer therapy. Trends Pharmacol Sci. 33:207–214. 2012. View Article : Google Scholar : PubMed/NCBI
3	Brown JM and Wilson WR: Exploiting tumour hypoxia in cancer treatment. Nat Rev Cancer. 4:437–447. 2004. View Article : Google Scholar : PubMed/NCBI
4	Birner P, Schindl M, Obermair A, Breitenecker G and Oberhuber G: Expression of hypoxia-inducible factor 1alpha in epithelial ovarian tumors: its impact on prognosis and on response to chemotherapy. Clin Cancer Res. 7:1661–1668. 2001.PubMed/NCBI
5	Rohwer N and Cramer T: Hypoxia-mediated drug resistance: novel insights on the functional interaction of HIFs and cell death pathways. Drug Resist Updat. 14:191–201. 2011. View Article : Google Scholar : PubMed/NCBI
6	Myzak MC, Dashwood WM, Orner GA, Ho E and Dashwood RH: Sulforaphane inhibits histone deacetylase in vivo and suppresses tumorigenesis in Apc-minus mice. FASEB J. 20:506–508. 2006.PubMed/NCBI
7	Chaudhuri D, Orsulic S and Ashok BT: Antiproliferative activity of sulforaphane in Akt-overexpressing ovarian cancer cells. Mol Cancer Ther. 6:334–345. 2007. View Article : Google Scholar : PubMed/NCBI
8	Bryant CS, Kumar S, Chamala S, Shah J, Pal J, Haider M, Seward S, Qazi AM, Morris R, Semaan A, et al: Sulforaphane induces cell cycle arrest by protecting RB-E2F-1 complex in epithelial ovarian cancer cells. Mol Cancer. 9:472010. View Article : Google Scholar : PubMed/NCBI
9	Chang CC, Hung CM, Yang YR, Lee MJ and Hsu YC: Sulforaphane induced cell cycle arrest in the G2/M phase via the blockade of cyclin B1/CDC2 in human ovarian cancer cells. J Ovarian Res. 6:412013. View Article : Google Scholar : PubMed/NCBI
10	Zhou C, Poulton EJ, Grün F, Bammler TK, Blumberg B, Thummel KE and Eaton DL: The dietary isothiocyanate sulforaphane is an antagonist of the human steroid and xenobiotic nuclear receptor. Mol Pharmacol. 71:220–229. 2007. View Article : Google Scholar
11	Bertl E, Bartsch H and Gerhäuser C: Inhibition of angiogenesis and endothelial cell functions are novel sulforaphane-mediated mechanisms in chemoprevention. Mol Cancer Ther. 5:575–585. 2006. View Article : Google Scholar : PubMed/NCBI
12	Yao H, Wang H, Zhang Z, Jiang BH, Luo J and Shi X: Sulforaphane inhibited expression of hypoxia-inducible factor-1alpha in human tongue squamous cancer cells and prostate cancer cells. Int J Cancer. 123:1255–1261. 2008. View Article : Google Scholar : PubMed/NCBI
13	Jeong JK, Moon MH, Seo JS, Seol JW, Lee YJ and Park SY: Sulforaphane blocks hypoxia-mediated resistance to TRAIL-induced tumor cell death. Mol Med Rep. 4:325–330. 2011.PubMed/NCBI
14	Kaelin WG Jr and Ratcliffe PJ: Oxygen sensing by metazoans: the central role of the HIF hydroxylase pathway. Mol Cell. 30:393–402. 2008. View Article : Google Scholar : PubMed/NCBI
15	Lendahl U, Lee KL, Yang H and Poellinger L: Generating specificity and diversity in the transcriptional response to hypoxia. Nat Rev Genet. 10:821–832. 2009. View Article : Google Scholar : PubMed/NCBI
16	Pijnenborg JM, Wijnakker M, Hagelstein J, Delvoux B and Groothuis PG: Hypoxia contributes to development of recurrent endometrial carcinoma. Int J Gynecol Cancer. 17:897–904. 2007. View Article : Google Scholar : PubMed/NCBI
17	Liang D, Ma Y, Liu J, Trope CG, Holm R, Nesland JM and Suo Z: The hypoxic microenvironment upgrades stem-like properties of ovarian cancer cells. BMC Cancer. 12:2012012. View Article : Google Scholar : PubMed/NCBI
18	Williams E, Martin S, Moss R, Durrant L and Deen S: Co-expression of VEGF and CA9 in ovarian high-grade serous carcinoma and relationship to survival. Virchows Arch. 461:33–39. 2012. View Article : Google Scholar : PubMed/NCBI
19	Sedlák J, Sedláková O, Hlavcák P, Hunáková L, Bízik J, Grófová M and Chorváth B: Cell surface phenotype and increased penetration of human multidrug-resistant ovarian carcinoma cells into in vitro collagen-fibroblasts matrix. Neoplasma. 43:389–395. 1996.PubMed/NCBI
20	Bodo J, Chovancova J, Hunakova L and Sedlak J: Enhanced sensitivity of human ovarian carcinoma cell lines A2780 and A2780/CP to the combination of cisplatin and synthetic isothiocyanate ethyl 4-isothiocyanatobutanoate. Neoplasma. 52:510–516. 2005.PubMed/NCBI
21	Svastová E, Hulíková A, Rafajová M, Zat’ovicová M, Gibadulinová A, Casini A, Cecchi A, Scozzafava A, Supuran CT, Pastorek J, et al: Hypoxia activates the capacity of tumor-associated carbonic anhydrase IX to acidify extracellular pH. FEBS Lett. 577:439–445. 2004. View Article : Google Scholar : PubMed/NCBI
22	Chen H, Landen CN, Li Y, Alvarez RD and Tollefsbol TO: Enhancement of cisplatin-mediated apoptosis in ovarian cancer cells through potentiating G2/M arrest and p21 upregulation by combinatorial epigallocatechin gallate and sulforaphane. J Oncol. 2013:8729572013. View Article : Google Scholar : PubMed/NCBI
23	Comerford KM, Cummins EP and Taylor CT: c-Jun NH2-terminal kinase activation contributes to hypoxia-inducible factor 1alpha-dependent P-glycoprotein expression in hypoxia. Cancer Res. 64:9057–9061. 2013. View Article : Google Scholar
24	An J, Liu H, Magyar CE, Guo Y, Veena MS, Srivatsan ES, Huang J and Rettig MB: Hyperactivated JNK is a therapeutic target in pVHL-deficient renal cell carcinoma. Cancer Res. 73:1374–1385. 2013. View Article : Google Scholar : PubMed/NCBI
25	Gillies RJ and Gatenby RA: Hypoxia and adaptive landscapes in the evolution of carcinogenesis. Cancer Metastasis Rev. 26:311–317. 2007. View Article : Google Scholar : PubMed/NCBI
26	Sibhatu MB, Smitherman PK, Townsend AJ and Morrow CS: Expression of MRP1 and GSTP1-1 modulate the acute cellular response to treatment with the chemopreventive isothiocyanate, sulforaphane. Carcinogenesis. 29:807–815. 2008. View Article : Google Scholar : PubMed/NCBI
27	Rudolf E, Andelová H and Cervinka M: Activation of several concurrent proapoptic pathways by sulforaphane in human colon cancer cells SW620. Food Chem Toxicol. 47:2366–2373. 2009. View Article : Google Scholar : PubMed/NCBI
28	Horiuchi A, Hayashi T, Kikuchi N, Hayashi A, Fuseya C, Shiozawa T and Konishi I: Hypoxia upregulates ovarian cancer invasiveness via the binding of HIF-1α to a hypoxia-induced, methylation-free hypoxia response element of S100A4 gene. Int J Cancer. 131:1755–1767. 2012. View Article : Google Scholar : PubMed/NCBI
29	Selvendiran K, Bratasz A, Kuppusamy ML, Tazi MF, Rivera BK and Kuppusamy P: Hypoxia induces chemoresistance in ovarian cancer cells by activation of signal transducer and activator of transcription 3. Int J Cancer. 125:2198–2204. 2009. View Article : Google Scholar : PubMed/NCBI
30	Milane L, Duan Z and Amiji M: Role of hypoxia and glycolysis in the development of multi-drug resistance in human tumor cells and the establishment of an orthotopic multi-drug resistant tumor model in nude mice using hypoxic pre-conditioning. Cancer Cell Int. 11:32011. View Article : Google Scholar : PubMed/NCBI
31	Shimogai R, Kigawa J, Itamochi H, Iba T, Kanamori Y, Oishi T, Shimada M, Sato S, Kawaguchi W, Sato S, et al: Expression of hypoxia-inducible factor 1alpha gene affects the outcome in patients with ovarian cancer. Int J Gynecol Cancer. 18:499–505. 2008. View Article : Google Scholar : PubMed/NCBI
32	Wong C, Wellman TL and Lounsbury KM: VEGF and HIF-1alpha expression are increased in advanced stages of epithelial ovarian cancer. Gynecol Oncol. 91:513–517. 2003. View Article : Google Scholar : PubMed/NCBI
33	Kim K, Park WY, Kim JY, Sol MY, Shin DH, Park do Y, Lee CH, Lee JH and Choi KU: Prognostic relevance of the expression of CA IX, GLUT-1, and VEGF in ovarian epithelial cancers. Korean J Pathol. 46:532–540. 2012. View Article : Google Scholar
34	Cheng JC, Klausen C and Leung PC: Hypoxia-inducible factor 1 alpha mediates epidermal growth factor-induced down-regulation of E-cadherin expression and cell invasion in human ovarian cancer cells. Cancer Lett. 329:197–206. 2013. View Article : Google Scholar
35	Hynninen P, Vaskivuo L, Saarnio J, Haapasalo H, Kivelä J, Pastoreková S, Pastorek J, Waheed A, Sly WS, Puistola U, et al: Expression of transmembrane carbonic anhydrases IX and XII in ovarian tumours. Histopathology. 49:594–602. 2006. View Article : Google Scholar : PubMed/NCBI
36	Jeon YK, Yoo DR, Jang YH, Jang SY and Nam MJ: Sulforaphane induces apoptosis in human hepatic cancer cells through inhibition of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase4, mediated by hypoxia inducible factor-1-dependent pathway. Biochim Biophys Acta. 1814.1340–1348. 2011.
37	Sermeus A and Michiels C: Reciprocal influence of the p53 and the hypoxic pathways. Cell Death Dis. 2:e1642011. View Article : Google Scholar : PubMed/NCBI
38	Chew YC, Adhikary G, Wilson GM, Xu W and Eckert RL: Sulforaphane induction of p21(Cip1) cyclin-dependent kinase inhibitor expression requires p53 and Sp1 transcription factors and is p53-dependent. J Biol Chem. 287:16168–16178. 2012. View Article : Google Scholar : PubMed/NCBI
39	Kolamunne RT, Dias IH, Vernallis AB, Grant MM and Griffiths HR: Nrf2 activation supports cell survival during hypoxia and hypoxia/reoxygenation in cardiomyoblasts; the roles of reactive oxygen and nitrogen species. Redox Biol. 1:418–426. 2013. View Article : Google Scholar : PubMed/NCBI
40	Kensler TW, Egner PA, Agyeman AS, Visvanathan K, Groopman JD, Chen JG, Chen TY, Fahey JW and Talalay P: Keap1-nrf2 signaling: a target for cancer prevention by sulforaphane. Top Curr Chem. 329:163–177. 2013. View Article : Google Scholar :
41	Chan WK, Yao G, Gu YZ and Bradfield CA: Cross-talk between the aryl hydrocarbon receptor and hypoxia inducible factor signaling pathways. Demonstration of competition and compensation. J Biol Chem. 274:12115–12123. 1999. View Article : Google Scholar : PubMed/NCBI
42	Takacova M, Holotnakova T, Vondracek J, Machala M, Pencikova K, Gradin K, Poellinger L, Pastorek J, Pastorekova S and Kopacek J: Role of aryl hydrocarbon receptor in modulation of the expression of the hypoxia marker carbonic anhydrase IX. Biochem J. 419:419–425. 2009. View Article : Google Scholar : PubMed/NCBI
43	Cavalli LR, Riggins RB, Wang A, Clarke R and Haddad BR: Frequent loss of heterozygosity at the interferon regulatory factor-1 gene locus in breast cancer. Breast Cancer Res Treat. 121:227–231. 2010. View Article : Google Scholar :
44	Wiczk A, Hofman D, Konopa G and Herman-Antosiewicz A: Sulforaphane, a cruciferous vegetable-derived isothiocyanate, inhibits protein synthesis in human prostate cancer cells. Biochim Biophys Acta. 1823.1295–1305. 2012.
45	Wykoff CC, Beasley NJ, Watson PH, Turner KJ, Pastorek J, Sibtain A, Wilson GD, Turley H, Talks KL, Maxwell PH, et al: Hypoxia-inducible expression of tumor-associated carbonic anhydrases. Cancer Res. 60:7075–7083. 2000.
46	Kaluz S, Kaluzová M, Opavský R, Pastoreková S, Gibadulinová A, Dequiedt F, Kettmann R and Pastorek J: Transcriptional regulation of the MN/CA 9 gene coding for the tumor-associated carbonic anhydrase IX. Identification and characterization of a proximal silencer element. J Biol Chem. 274:32588–32595. 1999. View Article : Google Scholar : PubMed/NCBI
47	Kaluzová M, Kaluz S, Lerman MI and Stanbridge EJ: DNA damage is a prerequisite for p53-mediated proteasomal degradation of HIF-1alpha in hypoxic cells and downregulation of the hypoxia marker carbonic anhydrase IX. Mol Cell Biol. 24:5757–5766. 2004. View Article : Google Scholar : PubMed/NCBI
48	Pastorek J, Pastoreková S, Callebaut I, Mornon JP, Zelník V, Opavský R, Zat’ovicová M, Liao S, Portetelle D, Stanbridge EJ, et al: Cloning and characterization of MN, a human tumor-associated protein with a domain homologous to carbonic anhydrase and a putative helix-loop-helix DNA binding segment. Oncogene. 9:2877–2888. 1994.PubMed/NCBI
49	Ditte P, Dequiedt F, Svastova E, Hulikova A, Ohradanova-Repic A, Zatovicova M, Csaderova L, Kopacek J, Supuran CT, Pastorekova S, et al: Phosphorylation of carbonic anhydrase IX controls its ability to mediate extracellular acidification in hypoxic tumors. Cancer Res. 71:7558–7567. 2011. View Article : Google Scholar : PubMed/NCBI
50	Fang JS, Gillies RD and Gatenby RA: Adaptation to hypoxia and acidosis in carcinogenesis and tumor progression. Semin Cancer Biol. 330–337. 2008. View Article : Google Scholar : PubMed/NCBI
51	Gatenby RA and Gillies RJ: A microenvironmental model of carcinogenesis. Nat Rev Cancer. 8:56–61. 2008. View Article : Google Scholar
52	Sedlakova O, Svastova E, Takacova M, Kopacek J, Pastorek J and Pastorekova S: Carbonic anhydrase IX, a hypoxia-induced catalytic component of the pH regulating machinery in tumors. Front Physiol. 4:4002014. View Article : Google Scholar : PubMed/NCBI
53	Jakubikova J, Cervi D, Ooi M, Kim K, Nahar S, Klippel S, Cholujova D, Leiba M, Daley JF, Delmore J, et al: Anti-tumor activity and signaling events triggered by the isothiocyanates, sulforaphane and phenethyl isothiocyanate, in multiple myeloma. Haematologica. 96:1170–1179. 2011. View Article : Google Scholar : PubMed/NCBI
54	Kaminski BM, Weigert A, Brüne B, Schumacher M, Wenzel U, Steinhilber D, Stein J and Ulrich S: Sulforaphane potentiates oxaliplatin-induced cell growth inhibition in colorectal cancer cells via induction of different modes of cell death. Cancer Chemother Pharmacol. 67:1167–1178. 2011. View Article : Google Scholar
55	Mokhtari RB, Kumar S, Islam SS, Yazdanpanah M, Adeli K, Cutz E and Yeger H: Combination of carbonic anhydrase inhibitor, acetazolamide, and sulforaphane, reduces the viability and growth of bronchial carcinoid cell lines. BMC Cancer. 13:3782013. View Article : Google Scholar : PubMed/NCBI