Triptolide induces autophagy and apoptosis through ERK activation in human breast cancer MCF‑7 cells

Gao,Huan; Zhang,Yue; Dong,Lei; Qu,Xiao‑Yu; Tao,Li‑Na; Zhang,Yue‑Ming; Zhai,Jing‑Hui; Song,Yan‑Qing

doi:10.3892/etm.2018.5830

Triptolide induces autophagy and apoptosis through ERK activation in human breast cancer MCF‑7 cells

Authors:
- Huan Gao
- Yue Zhang
- Lei Dong
- Xiao‑Yu Qu
- Li‑Na Tao
- Yue‑Ming Zhang
- Jing‑Hui Zhai
- Yan‑Qing Song
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Affiliations: Department of Pharmacy, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China
Published online on: February 1, 2018 https://doi.org/10.3892/etm.2018.5830
Pages: 3413-3419
Copyright: © Gao et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 4.0].

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Abstract

To investigate the effects of triptolide (TPI) on proliferation, autophagy and death in human breast cancer MCF‑7 cells, and to elucidate the associated molecular mechanisms, intracellular alterations were analyzed using 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT) and flow cytometry assays. The results of the MTT assay revealed that TPI significantly reduced the MCF‑7 cell survival rate when the concentration was >10 nmol/l. TPI activated a caspase cascade reaction by regulating Bcl‑2‑associated X protein (Bax), caspase‑3 and B‑cell lymphoma 2 expression, and promoted programmed cell death via the mitochondrial pathway. The results demonstrated that TPI significantly reduced the cell proliferation rate and viability in a time‑ and dose‑dependent manner, which was confirmed by western blotting and immunofluorescent staining. TPI induced autophagy and influenced p38 mitogen‑activated protein kinases, extracellular signal‑regulated kinase (Erk)1/2, and mammalian target of rapamycin (mTOR) phosphorylation, which resulted in apoptosis. When cells were treated with a combination of TPI and the Erk1/2 inhibitor U0126, the downregulation of P62 and upregulation of Bax were inhibited, which demonstrated that the inhibition of Erk1/2 reversed the autophagy changes induced by TPI. The results indicated that Erk1/2 activation may be a novel mechanism by which TPI induces autophagy and apoptosis in MCF‑7 breast cancer cells. In conclusion, TPI affects the proliferation and apoptosis of MCF‑7 cells, potentially via autophagy and p38/Erk/mTOR phosphorylation. The present study offers a novel view of the mechanisms by which TPI regulates cell death.

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1	Cedolini C, Bertozzi S, Londero AP, Bernardi S, Seriau L, Concina S, Cattin F and Risaliti A: Type of breast cancer diagnosis, screening, and survival. Clin Breast Cancer. 14:235–240. 2014. View Article : Google Scholar : PubMed/NCBI
2	de la Mare JA, Contu L, Hunter MC, Moyo B, Sterrenberg JN, Dhanani KC, Mutsvunguma LZ and Edkins AL: Breast cancer: Current developments in molecular approaches to diagnosis and treatment. Recent Pat Anticancer Drug Discov. 9:153–175. 2014. View Article : Google Scholar : PubMed/NCBI
3	Robijns J, Censabella S, Bulens P, Maes A and Mebis J: The use of low-level light therapy in supportive care for patients with breast cancer: Review of the literature. Lasers Med Sci. 32:229–242. 2017. View Article : Google Scholar : PubMed/NCBI
4	Ikeda H, Taira N, Nogami T, Shien K, Okada M, Shien T, Doihara H and Miyoshi S: Combination treatment with fulvestrant and various cytotoxic agents (doxorubicin, paclitaxel, docetaxel, vinorelbine and 5-fluorouracil) has a synergistic effect in estrogen receptor-positive breast cancer. Cancer Sci. 102:2038–2042. 2011. View Article : Google Scholar : PubMed/NCBI
5	Zheng RR, Hu W, Sui CG, Ma N and Jiang YH: Effects of doxorubicin and gemcitabine on the induction of apoptosis in breast cancer cells. Oncol Rep. 32:2719–2725. 2014. View Article : Google Scholar : PubMed/NCBI
6	Tacar O, Sriamornsak P and Dass CR: Doxorubicin: An update on anticancer molecular action, toxicity and novel drug delivery systems. J Pharm Pharmacol. 65:157–170. 2013. View Article : Google Scholar : PubMed/NCBI
7	Li XJ, Jiang ZZ and Zhang LY: Triptolide: Progress on research in pharmacodynamics and toxicology. J Ethnopharmacol. 155:67–79. 2014. View Article : Google Scholar : PubMed/NCBI
8	Reno TA, Kim JY and Raz DJ: Triptolide inhibits lung cancer cell migration, invasion, and metastasis. Ann Thorac Surg. 100:1817–1825. 2015. View Article : Google Scholar : PubMed/NCBI
9	Chen Z, Sangwan V, Banerjee S, Chugh R, Dudeja V, Vickers SM and Saluja AK: Triptolide sensitizes pancreatic cancer cells to TRAIL-induced activation of the death receptor pathway. Cancer Lett. 348:156–166. 2014. View Article : Google Scholar : PubMed/NCBI
10	Liu Y, Xiao E, Yuan L and Li G: Triptolide synergistically enhances antitumor activity of oxaliplatin in colon carcinoma in vitro and in vivo. DNA Cell Biol. 33:418–425. 2014. View Article : Google Scholar : PubMed/NCBI
11	Grossi V, Peserico A, Tezil T and Simone C: p38α MAPK pathway: A key factor in colorectal cancer therapy and chemoresistance. World J Gastroenterol. 20:9744–9758. 2014. View Article : Google Scholar : PubMed/NCBI
12	Zhou HF, Liu XY, Niu DB, Li FQ, He QH and Wang XM: Triptolide protects dopaminergic neurons from inflammation-mediated damage induced by lipopolysaccharide intranigral injection. Neurobiol Dis. 18:441–449. 2005. View Article : Google Scholar : PubMed/NCBI
13	Tang Y, Wang J, Cheng J and Wang L: Antiestrogenic activity of triptolide in human breast cancer cells MCF-7 and immature female mouse. Drug Dev Res. 78:164–169. 2017. View Article : Google Scholar : PubMed/NCBI
14	Liu J, Jiang Z, Xiao J, Zhang Y, Lin S, Duan W, Yao J, Liu C, Huang X, Wang T, et al: Effects of triptolide from Tripterygium wilfordii on ERalpha and p53 expression in two human breast cancer cell lines. Phytomedicine. 16:1006–1013. 2009. View Article : Google Scholar : PubMed/NCBI
15	Cheng X, Shi W, Zhao C, Zhang D, Liang P, Wang G and Lu L: Triptolide sensitizes human breast cancer cells to tumor necrosis factor-α-induced apoptosis by inhibiting activation of the nuclear factor-κB pathway. Mol Med Rep. 13:3257–3264. 2016. View Article : Google Scholar : PubMed/NCBI
16	Tan BJ and Chiu GN: Role of oxidative stress, endoplasmic reticulum stress and ERK activation in triptolide-induced apoptosis. Int J Oncol. 42:1605–1612. 2013. View Article : Google Scholar : PubMed/NCBI
17	Lewinska A, Adamczyk-Grochala J, Kwasniewicz E, Deregowska A and Wnuk M: Diosmin-induced senescence, apoptosis and autophagy in breast cancer cells of different p53 status and ERK activity. Toxicol Lett. 265:117–130. 2017. View Article : Google Scholar : PubMed/NCBI
18	Liu M, Chen J, Huang Y, Ke J, Li L, Huang D and Wu W: Triptolide alleviates isoprenaline-induced cardiac remodeling in rats via TGF-β1/Smad3 and p38 MAPK signaling pathway. Pharmazie. 70:244–250. 2015.PubMed/NCBI
19	Meng G, Wang W, Chai K, Yang S, Li F and Jiang K: Combination treatment with triptolide and hydroxycamptothecin synergistically enhances apoptosis in A549 lung adenocarcinoma cells through PP2A-regulated ERK, p38 MAPKs and Akt signaling pathways. Int J Oncol. 46:1007–1017. 2015. View Article : Google Scholar : PubMed/NCBI
20	Parzych KR and Klionsky DJ: An overview of autophagy: Morphology, mechanism, and regulation. Antioxid Redox Signal. 20:460–473. 2014. View Article : Google Scholar : PubMed/NCBI
21	Porter AG and Jänicke RU: Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 6:99–104. 1999. View Article : Google Scholar : PubMed/NCBI
22	Huang Q, Liu X, Cao C, Lei J, Han D, Chen G, Yu J, Chen L, Lv D and Li Z: Apelin-13 induces autophagy in hepatoma HepG2 cells through ERK1/2 signaling pathway-dependent upregulation of Beclin1. Oncol Lett. 11:1051–1056. 2016. View Article : Google Scholar : PubMed/NCBI
23	Koleini N and Kardami E: Autophagy and mitophagy in the context of doxorubicin-induced cardiotoxicity. Oncotarget. 8:46663–46680. 2017. View Article : Google Scholar : PubMed/NCBI
24	Zhao F, Huang W, Zhang Z, Mao L, Han Y, Yan J and Lei M: Triptolide induces protective autophagy through activation of the CaMKKβ-AMPK signaling pathway in prostate cancer cells. Oncotarget. 7:5366–5382. 2016.PubMed/NCBI
25	Li H, Pan GF, Jiang ZZ, Yang J, Sun LX and Zhang LY: Triptolide inhibits human breast cancer MCF-7 cell growth via downregulation of the ERα-mediated signaling pathway. Acta Pharmacol Sin. 36:606–613. 2015. View Article : Google Scholar : PubMed/NCBI
26	Jiang N, Dong XP, Zhang SL, You QY, Jiang XT and Zhao XG: Triptolide reverses the Taxol resistance of lung adenocarcinoma by inhibiting the NF-κB signaling pathway and the expression of NF-κB-regulated drug-resistant genes. Mol Med Rep. 13:153–159. 2016. View Article : Google Scholar : PubMed/NCBI
27	Kang DW, Lee JY, Oh DH, Park SY, Woo TM, Kim MK, Park MH, Jang YH and Min do S: Triptolide-induced suppression of phospholipase D expression inhibits proliferation of MDA-MB-231 breast cancer cells. Exp Mol Med. 41:678–685. 2009. View Article : Google Scholar : PubMed/NCBI
28	Kim YC and Guan KL: mTOR: A pharmacologic target for autophagy regulation. J Clin Invest. 125:25–32. 2015. View Article : Google Scholar : PubMed/NCBI
29	Meijer AJ, Lorin S, Blommaart EF and Codogno P: Regulation of autophagy by amino acids and MTOR-dependent signal transduction. Amino Acids. 47:2037–2063. 2015. View Article : Google Scholar : PubMed/NCBI
30	Shen P, Chen M, He M, Chen L, Song Y, Xiao P, Wan X, Dai F, Pan T and Wang Q: Inhibition of ERα/ERK/P62 cascades induces ‘autophagic switch’ in the estrogen receptor-positive breast cancer cells exposed to gemcitabine. Oncotarget. 7:48501–48516. 2016. View Article : Google Scholar : PubMed/NCBI
31	Yue X, Zhao P, Wu K, Huang J, Zhang W, Wu Y, Liang X and He X: GRIM-19 inhibition induced autophagy through activation of ERK and HIF-1α not STAT3 in Hela cells. Tumour Biol. 37:9789–9796. 2016. View Article : Google Scholar : PubMed/NCBI
32	Innajak S, Mahabusrakum W and Watanapokasin R: Goniothalamin induces apoptosis associated with autophagy activation through MAPK signaling in SK-BR-3 cells. Oncol Rep. 35:2851–2858. 2016. View Article : Google Scholar : PubMed/NCBI