Arctigenin inhibits the activation of the mTOR pathway, resulting in autophagic cell death and decreased ER expression in ER-positive human breast cancer cells

Maxwell,Thressi; Lee,Kyu  Shik; Kim,Soyoung; Nam,Kyung-Soo

doi:10.3892/ijo.2018.4271

Arctigenin inhibits the activation of the mTOR pathway, resulting in autophagic cell death and decreased ER expression in ER-positive human breast cancer cells

Authors:
- Thressi Maxwell
- Kyu Shik Lee
- Soyoung Kim
- Kyung-Soo Nam
View Affiliations

Affiliations: Department of Pharmacology, School of Medicine and Intractable Disease Research Center, Dongguk University, Gyeongju 38066, Republic of Korea
Published online on: February 9, 2018 https://doi.org/10.3892/ijo.2018.4271
Pages: 1339-1349

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Arctigenin, a member of the Asteraceae family, is a biologically active lignan that is consumed worldwide due to its several health benefits. However, its use may pose a problem for patients with estrogen receptor (ER)α-positive breast cancer, since studies have shown that arctigenin is a phytoestrogen that exerts a proliferative effect by binding to the ER. Thus, in this study, we examined the effect of arctigenin on ERα-positive MCF-7 human breast cancer cells to determine whether the consumption of arctigenin is safe for patients with breast cancer. First, we found that arctigenin inhibited the viability of the MCF-7 cells, and colony formation assay confirmed that this effect was cytotoxic rather than cytostatic. The cytotoxic effects were not mediated by cell cycle arrest, apoptosis, or necroptosis, despite DNA damage, as indicated by poly(ADP-ribose) polymerase (PARP) cleavage and phosphorylated H2A.X. An increase in lipidated LC3, a marker of autophagosome formation, was observed, indicating that autophagy was induced by arctigenin, which was found to be triggered by the inhibition of the mechanistic target of rapamycin (mTOR) pathway. We then examined the effects of arctigenin on ERα expression and determined whether it affects the sensitivity of the cells to tamoxifen, as tamoxifen is commonly used against hormone-responsive cancers and is known to act via the ERα. We found that treatment with arctigenin effectively downregulated ERα expression, which was found to be a consequence of the inhibition of the mTOR pathway. However, treatment with arctigenin in combination with tamoxifen did not affect the sensitivity of the cells to tamoxifen, but instead, exerted a synergistic effect. On the whole, our data indicate that the phytoestrogen, arctigenin, mainly targeted the mTOR pathway in ERα-positive MCF-7 human breast cancer cells, leading to autophagy-induced cell death and the downregulation of ERα expression. Furthermore, the synergistic effects between arctigenin and tamoxifen suggest that the consumption of arctigenin is not only safe for patients with hormone-sensitive cancers, but may also be an effective co-treatment.

View Figures

View References

1	Chan YS, Cheng LN, Wu JH, Chan E, Kwan YW, Lee SM, Leung GP, Yu PH and Chan SW: A review of the pharmacological effects of Arctium lappa (burdock). Inflammopharmacology. 19:245–254. 2011. View Article : Google Scholar
2	Hayashi K, Narutaki K, Nagaoka Y, Hayashi T and Uesato S: Therapeutic effect of arctiin and arctigenin in immunocompetent and immunocompromised mice infected with influenza A virus. Biol Pharm Bull. 33:1199–1205. 2010. View Article : Google Scholar : PubMed/NCBI
3	Swarup V, Ghosh J, Mishra MK and Basu A: Novel strategy for treatment of Japanese encephalitis using arctigenin, a plant lignan. J Antimicrob Chemother. 61:679–688. 2008. View Article : Google Scholar : PubMed/NCBI
4	Kim Y, Hollenbaugh JA, Kim DH and Kim B: Novel PI3K/Akt inhibitors screened by the cytoprotective function of human immunodeficiency virus type 1 Tat. PLoS One. 6:e217812011. View Article : Google Scholar : PubMed/NCBI
5	Kang HS, Lee JY and Kim CJ: Anti-inflammatory activity of arctigenin from Forsythiae Fructus. J Ethnopharmacol. 116:305–312. 2008. View Article : Google Scholar : PubMed/NCBI
6	Lee JY and Kim CJ: Arctigenin, a phenylpropanoid dibenzylbutyrolactone lignan, inhibits type I-IV allergic inflammation and pro-inflammatory enzymes. Arch Pharm Res. 33:947–957. 2010. View Article : Google Scholar : PubMed/NCBI
7	Zick SM, Sen A, Feng Y, Green J, Olatunde S and Boon H: Trial of Essiac to ascertain its effect in women with breast cancer (TEA-BC). J Altern Complement Med. 12:971–980. 2006. View Article : Google Scholar
8	Hsieh CJ, Kuo PL, Hsu YC, Huang YF, Tsai EM and Hsu YL: Arctigenin, a dietary phytoestrogen, induces apoptosis of estrogen receptor-negative breast cancer cells through the ROS/p38 MAPK pathway and epigenetic regulation. Free Radic Biol Med. 67:159–170. 2014. View Article : Google Scholar
9	Huang K, Li LA, Meng YG, You YQ, Fu XY and Song L: Arctigenin promotes apoptosis in ovarian cancer cells via the iNOS/NO/STAT3/survivin signalling. Basic Clin Pharmacol Toxicol. 115:507–511. 2014. View Article : Google Scholar : PubMed/NCBI
10	Jeong JB, Hong SC, Jeong HJ and Koo JS: Arctigenin induces cell cycle arrest by blocking the phosphorylation of Rb via the modulation of cell cycle regulatory proteins in human gastric cancer cells. Int Immunopharmacol. 11:1573–1577. 2011. View Article : Google Scholar : PubMed/NCBI
11	Jiang X, Zeng L, Huang J, Zhou H and Liu Y: Arctigenin, a natural lignan compound, induces apoptotic death of hepatocellular carcinoma cells via suppression of PI3-K/Akt signaling. J Biochem Mol Toxicol. 29:458–464. 2015. View Article : Google Scholar
12	Kim JY, Hwang JH, Cha MR, Yoon MY, Son ES, Tomida A, Ko B, Song SW, Shin-ya K, Hwang YI, et al: Arctigenin blocks the unfolded protein response and shows therapeutic antitumor activity. J Cell Physiol. 224:33–40. 2010.PubMed/NCBI
13	Susanti S, Iwasaki H, Inafuku M, Taira N and Oku H: Mechanism of arctigenin-mediated specific cytotoxicity against human lung adenocarcinoma cell lines. Phytomedicine. 21:39–46. 2013. View Article : Google Scholar : PubMed/NCBI
14	Wang HQ, Jin JJ and Wang J: Arctigenin enhances chemosensitivity to cisplatin in human nonsmall lung cancer H460 cells through downregulation of survivin expression. J Biochem Mol Toxicol. 28:39–45. 2014. View Article : Google Scholar : PubMed/NCBI
15	Wang L, Zhao F and Liu K: Induction of apoptosis of the human leukemia cells by arctigenin and its mechanism of action. Yao Xue Xue Bao. 43:542–547. 2008.In Chinese. PubMed/NCBI
16	Yang S, Ma J, Xiao J, Lv X, Li X, Yang H, Liu Y, Feng S and Zhang Y: Arctigenin anti-tumor activity in bladder cancer T24 cell line through induction of cell-cycle arrest and apoptosis. Anat Rec (Hoboken). 295:1260–1266. 2012. View Article : Google Scholar
17	Kang K, Lee HJ, Yoo JH, Jho EH, Kim CY, Kim M and Nho CW: Cell and nuclear enlargement of SW480 cells induced by a plant lignan, arctigenin: Evaluation of cellular DNA content using fluorescence microscopy and flow cytometry. DNA Cell Biol. 30:623–629. 2011. View Article : Google Scholar : PubMed/NCBI
18	Feng T, Cao W, Shen W, Zhang L, Gu X, Guo Y, Tsai HI, Liu X, Li J, Zhang J, et al: Arctigenin inhibits STAT3 and exhibits anticancer potential in human triple-negative breast cancer therapy. Oncotarget. 8:329–344. 2017.
19	Maxwell T, Chun SY, Lee KS, Kim S and Nam KS: The anti-metastatic effects of the phytoestrogen arctigenin on human breast cancer cell lines regardless of the status of ER expression. Int J Oncol. 50:727–735. 2017. View Article : Google Scholar
20	Wu X, Tong B, Yang Y, Luo J, Yuan X, Wei Z, Yue M, Xia Y and Dai Y: Arctigenin functions as a selective agonist of estrogen receptor β to restrict mTORC1 activation and consequent Th17 differentiation. Oncotarget. 7:83893–83906. 2016. View Article : Google Scholar : PubMed/NCBI
21	Bardin A, Boulle N, Lazennec G, Vignon F and Pujol P: Loss of ERbeta expression as a common step in estrogen-dependent tumor progression. Endocr Relat Cancer. 11:537–551. 2004. View Article : Google Scholar : PubMed/NCBI
22	Skliris GP, Munot K, Bell SM, Carder PJ, Lane S, Horgan K, Lansdown MR, Parkes AT, Hanby AM, Markham AF, et al: Reduced expression of oestrogen receptor beta in invasive breast cancer and its re-expression using DNA methyl transferase inhibitors in a cell line model. J Pathol. 201:213–220. 2003. View Article : Google Scholar : PubMed/NCBI
23	Xie LH, Ahn EM, Akao T, Abdel-Hafez AA, Nakamura N and Hattori M: Transformation of arctiin to estrogenic and antiestrogenic substances by human intestinal bacteria. Chem Pharm Bull (Tokyo). 51:378–384. 2003. View Article : Google Scholar
24	Chou TC and Talalay P: Quantitative analysis of dose-effect relationships: The combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul. 22:27–55. 1984. View Article : Google Scholar : PubMed/NCBI
25	Liang Y, Yan C and Schor NF: Apoptosis in the absence of caspase 3. Oncogene. 20:6570–6578. 2001. View Article : Google Scholar : PubMed/NCBI
26	Vandenabeele P, Galluzzi L, Vanden Berghe T and Kroemer G: Molecular mechanisms of necroptosis: An ordered cellular explosion. Nat Rev Mol Cell Biol. 11:700–714. 2010. View Article : Google Scholar : PubMed/NCBI
27	Wu X, Wu MY, Jiang M, Zhi Q, Bian X, Xu MD, Gong FR, Hou J, Tao M, Shou LM, et al: TNF-α sensitizes chemotherapy and radiotherapy against breast cancer cells. Cancer Cell Int. 17:132017. View Article : Google Scholar
28	Jung CH, Ro SH, Cao J, Otto NM and Kim DH: mTOR regulation of autophagy. FEBS Lett. 584:1287–1295. 2010. View Article : Google Scholar : PubMed/NCBI
29	Gu Y, Sun XX, Ye JM, He L, Yan SS, Zhang HH, Hu LH, Yuan JY and Yu Q: Arctigenin alleviates ER stress via activating AMPK. Acta Pharmacol Sin. 33:941–952. 2012. View Article : Google Scholar : PubMed/NCBI
30	Wu X, Dou Y, Yang Y, Bian D, Luo J, Tong B, Xia Y and Dai Y: Arctigenin exerts anti-colitis efficacy through inhibiting the differentiation of Th1 and Th17 cells via an mTORC1-dependent pathway. Biochem Pharmacol. 96:323–336. 2015. View Article : Google Scholar : PubMed/NCBI
31	Zhu Z, Yan J, Jiang W, Yao XG, Chen J, Chen L, Li C, Hu L, Jiang H and Shen X: Arctigenin effectively ameliorates memory impairment in Alzheimer's disease model mice targeting both β-amyloid production and clearance. J Neurosci. 33:13138–13149. 2013. View Article : Google Scholar : PubMed/NCBI
32	Yoshioka A, Miyata H, Doki Y, Yamasaki M, Sohma I, Gotoh K, Takiguchi S, Fujiwara Y, Uchiyama Y and Monden M: LC3, an autophagosome marker, is highly expressed in gastrointestinal cancers. Int J Oncol. 33:461–468. 2008.PubMed/NCBI
33	Chanoux RA, Yin B, Urtishak KA, Asare A, Bassing CH and Brown EJ: ATR and H2AX cooperate in maintaining genome stability under replication stress. J Biol Chem. 284:5994–6003. 2009. View Article : Google Scholar :
34	Koehn H, Magan N, Isaacs RJ and Stowell KM: Differential regulation of DNA repair protein Rad51 in human tumour cell lines exposed to doxorubicin. Anticancer Drugs. 18:419–425. 2007. View Article : Google Scholar : PubMed/NCBI
35	Maier P, Hartmann L, Wenz F and Herskind C: Cellular pathways in response to ionizing radiation and their targetability for tumor radiosensitization. Int J Mol Sci. 17:1–32. 2016. View Article : Google Scholar
36	Meijer AJ and Codogno P: Autophagy: Regulation by energy sensing. Curr Biol. 21:R227–R229. 2011. View Article : Google Scholar : PubMed/NCBI
37	Shagufta and Ahmad I: Tamoxifen a pioneering drug: An update on the therapeutic potential of tamoxifen derivatives. Eur J Med Chem. 143:515–531. 2018. View Article : Google Scholar
38	Lonard DM, Nawaz Z, Smith CL and O'Malley BW: The 26S proteasome is required for estrogen receptor-alpha and coactivator turnover and for efficient estrogen receptor-alpha transactivation. Mol Cell. 5:939–948. 2000. View Article : Google Scholar : PubMed/NCBI
39	Pinzone JJ, Stevenson H, Strobl JS and Berg PE: Molecular and cellular determinants of estrogen receptor alpha expression. Mol Cell Biol. 24:4605–4612. 2004. View Article : Google Scholar : PubMed/NCBI
40	Cui X, Zhang P, Deng W, Oesterreich S, Lu Y, Mills GB and Lee AV: Insulin-like growth factor-I inhibits progesterone receptor expression in breast cancer cells via the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin pathway: Progesterone receptor as a potential indicator of growth factor activity in breast cancer. Mol Endocrinol. 17:575–588. 2003. View Article : Google Scholar : PubMed/NCBI
41	Rice S and Whitehead SA: Phytoestrogens and breast cancer–promoters or protectors? Endocr Relat Cancer. 13:995–1015. 2006. View Article : Google Scholar : PubMed/NCBI
42	Matsumura A, Ghosh A, Pope GS and Darbre PD: Comparative study of oestrogenic properties of eight phytoestrogens in MCF7 human breast cancer cells. J Steroid Biochem Mol Biol. 94:431–443. 2005. View Article : Google Scholar : PubMed/NCBI
43	Lee J, Imm JY and Lee SH: β-catenin mediates anti-adipogenic and anticancer effects of arctigenin in preadipocytes and breast cancer cells. J Agric Food Chem. 65:2513–2520. 2017. View Article : Google Scholar : PubMed/NCBI
44	Lui A, New J, Ogony J, Thomas S and Lewis-Wambi J: Everolimus downregulates estrogen receptor and induces autophagy in aromatase inhibitor-resistant breast cancer cells. BMC Cancer. 16:4872016. View Article : Google Scholar : PubMed/NCBI
45	Nathan MR and Schmid P: A review of fulvestrant in breast cancer. Oncol Ther. 5:17–29. 2017. View Article : Google Scholar : PubMed/NCBI
46	Miller TW, Hennessy BT, González-Angulo AM, Fox EM, Mills GB, Chen H, Higham C, García-Echeverría C, Shyr Y and Arteaga CL: Hyperactivation of phosphatidylinositol-3 kinase promotes escape from hormone dependence in estrogen receptor-positive human breast cancer. J Clin Invest. 120:2406–2413. 2010. View Article : Google Scholar : PubMed/NCBI
47	Chumsri S, Sabnis G, Tkaczuk K and Brodie A: mTOR inhibitors: Changing landscape of endocrine-resistant breast cancer. Future Oncol. 10:443–456. 2014. View Article : Google Scholar : PubMed/NCBI
48	Ghayad SE, Bieche I, Vendrell JA, Keime C, Lidereau R, Dumontet C and Cohen PA: mTOR inhibition reverses acquired endocrine therapy resistance of breast cancer cells at the cell proliferation and gene-expression levels. Cancer Sci. 99:1992–2003. 2008.PubMed/NCBI