5,6,7,3',4',5'-Hexamethoxyflavone inhibits growth of triple-negative breast cancer cells via suppression of MAPK and Akt signaling pathways and arresting cell cycle

Borah,Natasha; Gunawardana,Shimara; Torres,Haydee; McDonnell,Susan; Van Slambrouck,Severine

doi:10.3892/ijo.2017.4157

5,6,7,3',4',5'-Hexamethoxyflavone inhibits growth of triple-negative breast cancer cells via suppression of MAPK and Akt signaling pathways and arresting cell cycle

Authors:
- Natasha Borah
- Shimara Gunawardana
- Haydee Torres
- Susan McDonnell
- Severine Van Slambrouck
View Affiliations

Affiliations: Department of Chemistry and Biochemistry, South Dakota State University, Brookings, SD 57007, USA, UCD School of Chemical and Bioprocess Engineering, University College Dublin, Belfield Dublin 4, Ireland
Published online on: October 13, 2017 https://doi.org/10.3892/ijo.2017.4157
Pages: 1685-1693
Copyright: © Borah et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Natural components continue to be an important source for the discovery and development of novel anticancer agents. Polymethoxyflavones are a class of flavonoids found in citrus fruits and medicinal plants used in traditional medicine. In the present study, the anticancer activity of the well-known nobiletin (5,6,7,8,3',4'-hexamethoxyflavone) was compared against its less studied structural isomer 5,6,7,3',4',5'-hexamethoxyflavone. These compounds were evaluated on the Hs578T triple-negative breast cancer cell line and its more migratory subclone Hs578Ts(i)8. 5,6,7,3',4',5'-hexamethoxyflavone was found to be less toxic than nobiletin, while a similar growth inhibitory effect was observed after 72 h. Additionally, 5,6,7,3',4',5'-hexamethoxyflavone arrested the cell cycle in the G2/M phase, while no effect was observed on apoptosis or the migratory behavior of these cells. Furthermore, mechanistic studies revealed that the growth inhibition was concomitant with reduced phosphorylation levels of signaling molecules in the MAPK and Akt pathways as well as cell cycle regulators, involved in regulating cell proliferation, survival and cell cycle. In summary, the present study is the first to report on the anticancer activities of 5,6,7,3',4',5'-hexamethoxyflavone and to provide evidence that this flavone could have a greater potential than nobiletin for prevention or treatment of triple- negative breast cancer.

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1	Newman DJ and Cragg GM: Natural products as sources of new drugs over the last 25 years. J Nat Prod. 70:461–477. 2007. View Article : Google Scholar : PubMed/NCBI
2	Orphan Drug status for alvocidib. Oncology Times. 25–May. 2014, https://doi.org/10.1097/01.COT.0000450374.31680.1d.
3	Sak K: Cytotoxicity of dietary flavonoids on different human cancer types. Pharmacogn Rev. 8:122–146. 2014. View Article : Google Scholar : PubMed/NCBI
4	Walle T: Methoxylated flavones, a superior cancer chemopreventive flavonoid subclass? Semin Cancer Biol. 17:354–362. 2007. View Article : Google Scholar : PubMed/NCBI
5	Mizuno M, Iinuma M, Ohara M, Tanaka T and Iwamasa M: Chemotaxonomy of the genus Citrus based on polymethoxyflavones. Chem Pharm Bull (Tokyo). 39:945–949. 1991. View Article : Google Scholar
6	Yenjai C, Prasanphen K, Daodee S, Wongpanich V and Kittakoop P: Bioactive flavonoids from Kaempferia parviflora. Fitoterapia. 75:89–92. 2004. View Article : Google Scholar
7	Faqueti LG, Brieudes V, Halabalaki M, Skaltsounis AL, Nascimento LF, Barros WM, Santos AR and Biavatti MW: Antinociceptive and anti-inflammatory activities of standardized extract of polymethoxyflavones from Ageratum conyzoides. J Ethnopharmacol. 194:369–377. 2016. View Article : Google Scholar : PubMed/NCBI
8	Lien LM, Wang MJ, Chen RJ, Chiu HC, Wu JL, Shen MY, Chou DS, Sheu JR, Lin KH and Lu WJ: Nobiletin, a polymethoxylated flavone, inhibits glioma cell growth and migration via arresting cell cycle and suppressing MAPK and Akt pathways. Phytother Res. 30:214–221. 2016. View Article : Google Scholar
9	Huang H, Li L, Shi W, Liu H, Yang J, Yuan X and Wu L: The multifunctional effects of nobiletin and its metabolites in vivo and in vitro. Evid Based Complement Alternat Med. 2016:29187962016. View Article : Google Scholar :
10	Chien SY, Hsieh MJ, Chen CJ, Yang SF and Chen MK: Nobiletin inhibits invasion and migration of human nasopharyngeal carcinoma cell lines by involving ERK1/2 and transcriptional inhibition of MMP-2. Expert Opin Ther Targets. 19:307–320. 2015. View Article : Google Scholar : PubMed/NCBI
11	Surichan S, Androutsopoulos VP, Sifakis S, Koutala E, Tsatsakis A, Arroo RR and Boarder MR: Bioactivation of the citrus flavonoid nobiletin by CYP1 enzymes in MCF7 breast adenocarcinoma cells. Food Chem Toxicol. 50:3320–3328. 2012. View Article : Google Scholar : PubMed/NCBI
12	Chen C, Ono M, Takeshima M and Nakano S: Antiproliferative and apoptosis-inducing activity of nobiletin against three subtypes of human breast cancer cell lines. Anticancer Res. 34:1785–1792. 2014.PubMed/NCBI
13	Andreopoulou E, Schweber SJ, Sparano JA and McDaid HM: Therapies for triple negative breast cancer. Expert Opin Pharmacother. 16:983–998. 2015. View Article : Google Scholar : PubMed/NCBI
14	Adebayo AH, Jig CJ, Zhang YM, He WJ, Zeng GZ, Han HJ, Xu JJ, Akindahunsi AA and Tana NH: A new chromene isolated from Ageratum conyzoides. Nat Prod Commun. 6:1263–1265. 2011.PubMed/NCBI
15	Itoh T, Ohguchi K, Iinuma M, Nozawa Y and Akao Y: Inhibitory effects of polymethoxy flavones isolated from Citrus reticulate on degranulation in rat basophilic leukemia RBL-2H3: Enhanced inhibition by their combination. Bioorg Med Chem. 16:7592–7598. 2008. View Article : Google Scholar : PubMed/NCBI
16	Hughes L, Malone C, Chumsri S, Burger AM and McDonnell S: Characterisation of breast cancer cell lines and establishment of a novel isogenic subclone to study migration, invasion and tumourigenicity. Clin Exp Metastasis. 25:549–557. 2008. View Article : Google Scholar : PubMed/NCBI
17	Romijn JC, Verkoelen CF and Schroeder FH: Application of the MTT assay to human prostate cancer cell lines in vitro: Establishment of test conditions and assessment of hormone-stimulated growth and drug-induced cytostatic and cytotoxic effects. Prostate. 12:99–110. 1988. View Article : Google Scholar : PubMed/NCBI
18	Rasband WS: ImageJ. U. S. National Institutes of Health; Bethesda, Maryland, USA: https://imagej.nih.gov/ij/. 1997–2016
19	Payan I, McDonnell S, Torres HM, Steelant WF and Van Slambrouck S: FAK tyrosine 407 organized with integrin αVβ5 in Hs578Ts(i)₈ advanced triple-negative breast cancer cells. Int J Oncol. 48:2043–2054. 2016. View Article : Google Scholar : PubMed/NCBI
20	Mustacchi G and De Laurentiis M: The role of taxanes in triple-negative breast cancer: Literature review. Drug Des Devel Ther. 9:4303–4318. 2015. View Article : Google Scholar : PubMed/NCBI
21	Lehmann BD, Bauer JA, Chen X, Sanders ME, Chakravarthy AB, Shyr Y and Pietenpol JA: Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. J Clin Invest. 121:2750–2767. 2011. View Article : Google Scholar : PubMed/NCBI
22	Van Slambrouck S, Parmar VS, Sharma SK, De Bondt B, Foré F, Coopman P, Vanhoecke BW, Boterberg T, Depypere HT, Leclercq G, et al: Tangeretin inhibits extracellular-signal-regulated kinase (ERK) phosphorylation. FEBS Lett. 579:1665–1669. 2005. View Article : Google Scholar : PubMed/NCBI
23	McConathy J and Owens MJ: Stereochemistry in drug action. Prim Care Companion J Clin Psychiatry. 5:70–73. 2003. View Article : Google Scholar
24	Baek SH, Kim SM, Nam D, Lee JH and Ahn KS, Choi SH, Kim SH, Shim BS, Chang IM and Ahn KS: Antimetastatic effect of nobiletin through the down-regulation of CXC chemokine receptor type 4 and matrix metallopeptidase-9. Pharm Biol. 50:1210–1218. 2012. View Article : Google Scholar : PubMed/NCBI
25	Sp N, Kang DY, Joung YH, Park JH, Kim WS, Lee HK, Song KD, Park YM and Yang YM: Nobiletin inhibits angiogenesis by regulating Src/FAK/STAT3-mediated signaling through PXN in ER⁺ breast cancer cells. Int J Mol Sci. 18:9352017. View Article : Google Scholar
26	Dong Y, Ji G, Cao A, Shi J, Shi H, Xie J and Wu D: Effects of sinensetin on proliferation and apoptosis of human gastric cancer AGS cells. Zhongguo Zhong Yao Za Zhi. 36:790–794. 2011.In Chinese. PubMed/NCBI
27	Burkhard K and Shapiro P: Use of inhibitors in the study of MAP kinases. Methods Mol Biol. 661:107–122. 2010. View Article : Google Scholar : PubMed/NCBI
28	Zannini L, Delia D and Buscemi G: CHK2 kinase in the DNA damage response and beyond. J Mol Cell Biol. 6:442–457. 2014. View Article : Google Scholar : PubMed/NCBI
29	Singh SV, Herman-Antosiewicz A, Singh AV, Lew KL, Srivastava SK, Kamath R, Brown KD, Zhang L and Baskaran R: Sulforaphane-induced G2/M phase cell cycle arrest involves checkpoint kinase 2-mediated phosphorylation of cell division cycle 25C. J Biol Chem. 279:25813–25822. 2004. View Article : Google Scholar : PubMed/NCBI
30	Sancar A, Lindsey-Boltz LA, Unsal-Kaçmaz K and Linn S: Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu Rev Biochem. 73:39–85. 2004. View Article : Google Scholar : PubMed/NCBI
31	Zhou BB and Bartek J: Targeting the checkpoint kinases: Chemosensitization versus chemoprotection. Nat Rev Cancer. 4:216–225. 2004. View Article : Google Scholar : PubMed/NCBI
32	DiPaola RS: To arrest or not to G2-M cell-cycle arrest. Clin Cancer Res. 8:3512–3519. 2002.