Potent antitumor activity of cepharanthine against triple‑negative breast cancer spheroids compared with tetrandrine

Kiyomi,Anna; Miyakawa,Risako; Matsumoto,Juri; Yamazaki,Kyousuke; Imai,Shinobu; Yuan,Bo; Hirano,Toshihiko; Sugiura,Munetoshi

doi:10.3892/ol.2020.12191

Potent antitumor activity of cepharanthine against triple‑negative breast cancer spheroids compared with tetrandrine

Authors:
- Anna Kiyomi
- Risako Miyakawa
- Juri Matsumoto
- Kyousuke Yamazaki
- Shinobu Imai
- Bo Yuan
- Toshihiko Hirano
- Munetoshi Sugiura
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Affiliations: Department of Drug Safety and Risk Management, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192‑0392, Japan, Laboratory of Pharmacology, School of Pharmacy, Josai University, Sakado, Saitama 350‑0295, Japan, Department of Clinical Pharmacology, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo 192‑0392, Japan
Published online on: October 6, 2020 https://doi.org/10.3892/ol.2020.12191
Article Number: 331
Copyright: © Kiyomi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cepharanthine (CEP) is a bis‑bynzelisoquinoline alkaloid from the same class as the anticancer agent tetrandrine (TET). However, the effects of CEP against breast cancer have not been extensively studied, despite its long therapeutic history with low toxicity against other types of cancer. 3D culture systems more accurately mimic the human body and address the limitations of determining drug effectiveness compared with 2D culture systems. In the present study, the antitumor activities of TET and CEP were compared in 3D culture systems in triple‑negative breast cancer (TNBC) MDA‑MB‑231 and estrogen receptor‑positive breast cancer MCF‑7 cell lines. Cell viability, apoptosis and cytotoxicity assays were performed to determine the total number of live or dead cells, the IC₅₀ values, the number of apoptotic cells and spheroid roundness. Viability suppression of MDA‑MB‑231 cells was significantly greater with both TET and CEP compared with that of MCF‑7 cells, and the roundness of MDA‑MB‑231 spheroids treated with CEP was decreased significantly compared with that of spheroid treated with TET. Cytoplasmic shrinkage in each cell line significantly increased with the treatment of TET compared with the control; however, this effect was stronger with CEP. The ratio of dead/live cells in each cell line treated with TET and CEP increased in a dose‑dependent manner. Overall, the present study demonstrated that CEP had greater cell toxicity in 3D spheroids of breast cancer cells compared with TET, suggesting that CEP may have a stronger antitumor activity on TNBC spheroids compared with TET.

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1	Tao Z, Shi A, Lu C, Song T, Zhang Z and Zhao J: Breast cancer: Epidemiology and etiology. Cell Biochem Biophys. 72:333–338. 2015. View Article : Google Scholar : PubMed/NCBI
2	Coughlin S: Epidemiology of breast cancer in women. Adv Exp Med Biol. 1152:9–29. 2019. View Article : Google Scholar : PubMed/NCBI
3	Winters S, Martin C, Murphy D and Shokar N: Breast cancer epidemiology, prevention, and screening. Prog Mol Biol Transl Sci. 151:1–32. 2017. View Article : Google Scholar : PubMed/NCBI
4	Harbeck N and Gnant M: Breast cancer. Lancet. 389:1134–1150. 2017. View Article : Google Scholar : PubMed/NCBI
5	Carey L, Winer E, Viale G, Cameron D and Gianni L: Triple-negative breast cancer: Disease entity or title of convenience? Nat Rev Clin Oncol. 7:683–692. 2010. View Article : Google Scholar : PubMed/NCBI
6	Liedtke C, Mazouni C, Hess K, André F, Tordai A, Mejia J, Symmans WF, Gonzalez-Angulo A, Hennessy B, Green M, et al: Response to neoadjuvant therapy and long-term survival in patients with triple-negative breast cancer. J Clin Oncol. 26:1275–1281. 2008. View Article : Google Scholar : PubMed/NCBI
7	Rakha EA, El-Sayed ME, Green AR, Lee AH, Robertson JF and Ellis IO: Prognostic markers in triple-negative breast cancer. Cancer. 109:25–32. 2007. View Article : Google Scholar : PubMed/NCBI
8	Denkert C, Liedtke C, Tutt A and von Minckwitz G: Molecular alterations in triple-negative breast cancer-the road to new treatment strategies. Lancet. 389:2430–2442. 2017. View Article : Google Scholar : PubMed/NCBI
9	Thomford NE, Senthebane DA, Rowe A, Munro D, Seele P, Maroyi A and Dzobo K: Natural products for drug discovery in the 21st century: Innovations for novel drug discovery. Int J Mol Sci. 19:15782018. View Article : Google Scholar
10	Harvey AL: Natural products in drug discovery. Drug Discov Today. 13:894–901. 2008. View Article : Google Scholar : PubMed/NCBI
11	Chen Y: Potential role of tetrandrine in cancer therapy. Acta Pharmacol Sin. 23:1102–1106. 2002.PubMed/NCBI
12	Liu T, Liu X and Li W: Tetrandrine, a Chinese plant-derived alkaloid, is a potential candidate for cancer chemotherapy. Oncotarget. 7:40800–40815. 2016. View Article : Google Scholar : PubMed/NCBI
13	Liu B, Wang T, Qian X, Liu G, Yu L and Ding Y: Anticancer effect of tetrandrine on primary cancer cells isolated from ascites and pleural fluids. Cancer Lett. 268:166–175. 2008. View Article : Google Scholar : PubMed/NCBI
14	Lu Y, Li F, Xu T and Sun J: Tetrandrine prevents multidrug resistance in the osteosarcoma cell line, U-2OS, by preventing Pgp overexpression through the inhibition of NF-κB signaling. Int J Mol Med. 39:993–1000. 2017. View Article : Google Scholar : PubMed/NCBI
15	Zhu R, Liu T, Tan Z, Wu X, Li M, Jiang L, Bao R, Shu Y, Lu A and Liu Y: Tetrandrine induces apoptosis in gallbladder carcinoma in vitro. Int J Clin Pharmacol Ther. 52:900–905. 2014. View Article : Google Scholar : PubMed/NCBI
16	Sun XC, Cheng HY, Deng YX, Shao RG and Ma J: Tetrandrine: A potent abrogator of G2 checkpoint function in tumor cells and its mechanism. Biomed Environ Sci. 20:495–501. 2007.PubMed/NCBI
17	Qiu W, Zhang AL and Tian Y: Tetrandrine triggers an alternative autophagy in DU145 cells. Oncol Lett. 13:3734–3738. 2017. View Article : Google Scholar : PubMed/NCBI
18	Xu WL, Shen HL, Ao ZF, Chen BA, Xia W, Gao F and Zhang YN: Combination of tetrandrine as a potential-reversing agent with daunorubicin, etoposide and cytarabine for the treatment of refractory and relapsed acute myelogenous leukemia. Leuk Res. 30:407–413. 2006. View Article : Google Scholar : PubMed/NCBI
19	Liu W, Zhang J, Ying C, Wang Q, Yan C, Jingyue Y, Zhaocai Y, Yan X, Heng-jun S and Lin J: Tetrandrine combined with gemcitabine and cisplatin for patients with advanced non-small cell lung cancer improve efficacy. Int J Biomed Sci. 8:28–35. 2012.PubMed/NCBI
20	Bailly C: Cepharanthine: An update of its mode of action, pharmacological properties and medical applications. Phytomedicine. 62:1529562019. View Article : Google Scholar : PubMed/NCBI
21	Asaumi J, Nishikawa K, Matsuoka H, Iwata M, Kawasaki S, Hiraki Y and Nishijima K: Direct antitumor effect of cepharanthin and combined effect with adriamycin against Ehrlich ascites tumor in mice. Anticancer Res. 15:67–70. 1995.PubMed/NCBI
22	Ono M and Tanaka N: Positive interaction of bisbenzylisoquinoline alkaloid, cepharanthin, with vinca alkaloid agents against human tumors. In Vivo. 11:233–241. 1997.PubMed/NCBI
23	Rosso F, Giordano A, Barbarisi M and Barbarisi A: From cell-ECM interactions to tissue engineering. J Cell Physiol. 199:174–180. 2004. View Article : Google Scholar : PubMed/NCBI
24	Ryu N, Lee S and Park H: Spheroid culture system methods and applications for mesenchymal stem cells. Cells. 8:16202019. View Article : Google Scholar
25	Breslin S and O'Driscoll L: The relevance of using 3D cell cultures, in addition to 2D monolayer cultures, when evaluating breast cancer drug sensitivity and resistance. Oncotarget. 7:45745–45756. 2016. View Article : Google Scholar : PubMed/NCBI
26	Imamura Y, Mukohara T, Shimono Y, Funakoshi Y, Chayahara N, Toyoda M, Kiyota N, Takao S, Kono S, Nakatsura T and Minami H: Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer. Oncol Rep. 33:1837–1843. 2015. View Article : Google Scholar : PubMed/NCBI
27	Moscona A: Rotation-mediated histogenetic aggregation of dissociated cells. A quantifiable approach to cell interactions in vitro. Exp Cell Res. 22:455–475. 1961. View Article : Google Scholar : PubMed/NCBI
28	O'Keane JC, Kupchik HZ, Schroy PC, Andry CD, Collins E and O'Brien MJ: A three-dimensional system for long-term culture of human colorectal adenomas. Am J Pathol. 137:1539–1547. 1990.PubMed/NCBI
29	Carlsson J, Nilsson K, Westermark B, Pontén J, Sundström C, Larsson E, Bergh J, Påhlman S, Busch C and Collins VP: Formation and growth of multicellular spheroids of human origin. Int J Cancer. 31:523–533. 1983. View Article : Google Scholar : PubMed/NCBI
30	Kleinman HK, McGarvey ML, Hassell JR, Star VL, Cannon FB, Laurie GW and Martin GR: Basement membrane complexes with biological activity. Biochemistry. 25:312–318. 1986. View Article : Google Scholar : PubMed/NCBI
31	Lawler EM, Miller FR and Heppner GH: Significance of three-dimensional growth patterns of mammary tissues in collagen gels. In Vitro. 19:600–610. 1983. View Article : Google Scholar : PubMed/NCBI
32	Bresciani G, Hofland LJ, Dogan F, Giamas G, Gagliano T and Zatelli MC: Evaluation of spheroid 3D culture methods to study a pancreatic neuroendocrine neoplasm cell line. Front Endocrinol (Lausanne). 10:6822019. View Article : Google Scholar : PubMed/NCBI
33	Nath S and Devi G: Three-dimensional culture systems in cancer research: Focus on tumor spheroid model. Pharmacol Ther. 163:94–108. 2016. View Article : Google Scholar : PubMed/NCBI
34	Kelm JM, Timmins NE, Brown CJ, Fussenegger M and Nielsen LK: Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. Biotechnol Bioeng. 83:173–180. 2003. View Article : Google Scholar : PubMed/NCBI
35	Vinci M, Gowan S, Boxall F, Patterson L, Zimmermann M, Court W, Lomas C, Mendiola M, Hardisson D and Eccles SA: Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation. BMC Biol. 10:292012. View Article : Google Scholar : PubMed/NCBI
36	Xu W, Wang X, Tu Y, Masaki H, Tanaka S, Onda K, Sugiyama K, Yamada H and Hirano T: Tetrandrine and cepharanthine induce apoptosis through caspase cascade regulation, cell cycle arrest, MAPK activation and PI3K/Akt/mTOR signal modification in glucocorticoid resistant human leukemia Jurkat T cells. Chem Biol Interact. 130:1087262019. View Article : Google Scholar
37	Nigjeh SE, Yeap SK, Nordin N, Kamalideghan B, Ky H and Rosli R: Citral induced apoptosis in MDA-MB-231 spheroid cells. BMC Complement Altern Med. 18:562018. View Article : Google Scholar : PubMed/NCBI
38	Jhan JR and Andrechek ER: Triple-negative breast cancer and the potential for targeted therapy. Pharmacogenomics. 18:1595–1609. 2017. View Article : Google Scholar : PubMed/NCBI
39	Bhagya N and Chandrashekar KR: Tetrandrine-A molecule of wide bioactivity. Phytochemistry. 125:5–13. 2016. View Article : Google Scholar : PubMed/NCBI
40	Gatti R, Belletti S, Orlandini G, Bussolati O, Dall'Asta V and Gazzola GC: Comparison of annexin V and calcein-AM as early vital markers of apoptosis in adherent cells by confocal laser microscopy. J Histochem Cytochem. 46:895–900. 1998. View Article : Google Scholar : PubMed/NCBI
41	Basmaciyan L, Azas N and Casanova M: Calcein+/PI-as an early apoptotic feature in Leishmania. PLoS One. 12:e01877562017. View Article : Google Scholar : PubMed/NCBI
42	Leung YM, Berdik M, Kwan CY and Loh TT: Effects of tetrandrine and closely related bis-benzylisoquinoline derivatives on cytosolic Ca²⁺ in human leukemic HL-60 cells: A structure-activity relationship study. Clin Exp Pharmacol Physiol. 23:653–659. 1996. View Article : Google Scholar : PubMed/NCBI
43	Kunz-Schughart L, Freyer J, Hofstaedter F and Ebner R: The use of 3-D cultures for high-throughput screening: The multicellular spheroid model. J Biomol Screen. 9:273–285. 2004. View Article : Google Scholar : PubMed/NCBI
44	Kiyomi A, Makita M, Ozeki T, Li N, Satomura A, Tanaka S, Onda K, Sugiyama K, Iwase T and Hirano T: Characterization and clinical implication of Th1/Th2/Th17 cytokines produced from three-dimensionally cultured tumor tissues resected from breast cancer patients. Transl Oncol. 8:318–326. 2015. View Article : Google Scholar : PubMed/NCBI
45	Jo Y, Choi N, Kim K, Koo HJ, Choi J and Kim HN: Chemoresistance of cancer cells: Requirements of tumor microenvironment-mimicking in vitro models in anti-cancer drug development. Theranostics. 8:5259–5275. 2018. View Article : Google Scholar : PubMed/NCBI
46	Zhou P, Li Z, Xu D, Wang Y, Bai Q, Feng Y, Su G, Chen P, Wang Y, Liu H, et al: Cepharanthine hydrochloride improves cisplatin chemotherapy and enhances immunity by regulating intestinal microbes in mice. Front Cell Infect Microbiol. 9:2252019. View Article : Google Scholar : PubMed/NCBI