Niclosamide Suppresses Proliferation, Induces Apoptosis and Inhibits Wnt/β-catenin Signaling Pathway in Human Ovarian Cancer Cells

  • Ayşe Çakır Gündoğdu Gazi University
  • Gülnur TAKE KAPLANOĞLU Gazi University
  • Hülya Sivas Anadolu University
  • Reyhan Varol Anadolu University
  • Cemile Merve Seymen Gazi University
Keywords: OVCAR-3, niclosamide, 5-fluorouracil, Wnt/β-catenin, proliferation, apoptosis

Abstract

Objective: The aim of this study is to investigate in vitro effects of an antihelminthic drug niclosamide on human ovarian carcinoma cell line OVCAR-3. Methods: MTT assay was applied to investigate the cytotoxic effects of niclosamide on the cells. β-catenin levels in the cells were analyzed by immunocytochemistry, in order to assess the potency of niclosamide on Wnt/β-catenin signaling pathway that function in cell proliferation. The effects of the drug on apoptosis were detected by TUNEL method. All the assays were also performed for chemotherapy agent 5-fluorouracil (5-FU) and anticancer effects of these two drugs were compared. Results: It was found that niclosamide at 1 μM and 2 μM concentrations reduced cell viability, whereas 5-FU showed its significant proliferation inhibitory effect at higher concentrations. Niclosamide led to an increase in apoptosis while this effect was weaker compared with 5-FU. Niclosamide treatment decreased β-catenin staining in the cells significantly but 5-FU did not affect β-catenin levels. Conclusion: The results indicate that niclosamide induces apoptosis and suppresses cell proliferation by inhibiting Wnt/β-catenin signaling pathway in OVCAR-3 cells. In conclusion, these findings warrant further evaluation of niclosamide as a promising therapy for ovarian cancer.

Author Biographies

Ayşe Çakır Gündoğdu, Gazi University
Department of Histology and Embryology, Faculty of Medicine, Gazi University, Ankara, Turkey M.Sc.
Gülnur TAKE KAPLANOĞLU, Gazi University
Department of Histology and Embryology, Faculty of Medicine, Gazi University, Ankara, Turkey Ph.D.
Hülya Sivas, Anadolu University
Department of Biology, Faculty of Science, Anadolu University, Eskisehir, Turkey Ph.D.
Reyhan Varol, Anadolu University
Department of Biology, Faculty of Science, Anadolu University, Eskisehir, Turkey
Cemile Merve Seymen, Gazi University
Department of Histology and Embryology, Faculty of Medicine, Gazi University, Ankara, Turkey

References

Lowe KA, Chia VM, Taylor A, O’Malley C, Kelsh M, Mohamed M, et al. An international assessment of ovarian cancer incidence and mortality. Gynecol Oncol 2013; 130:107-114.

Vargas-Hernández VM, Moreno-Eutimio MA, Acosta-Altamirano G, Vargas-Aguilar VM. Management of recurrent epithelial ovarian cancer. Gland Surg 2014; 3:198-202.

Garin J, Despeignes J, Billerau M: Present treatment of taeniasis with niclosamide. Lyon Med 1964; 212:1581-8.

Pearson RD, Hewlett EL. Niclosamide therapy for tapeworm infections. Ann Intern Med 1985; 102:550-1.

Weinbach EC, Garbus J. Mechanism of action of reagents that uncouple oxidative phosphorylation. Nature 1969; 221:1016–8.

Li R, You S, Hu Z, Chen ZG, Sica GL, Khuri FR, et al. Inhibition of STAT3 by niclosamide synergizes with erlotinib against head and neck cancer. PloS one 2013; 8: e74670.

Shi L, Zheng H, Hu W, Zhou B, Dai X, Zhang Y, Liu Z, Wu X, Zhao C, Liang G. Niclosamide inhibition of STAT3 synergizes with erlotinib in human colon cancer. Onco Targets Ther 2017; 10:1767-776.

Gangrade A, Pathak V, Augelli-Szafran CE, Wei HX, Oliver P, Suto M, et al. Preferential Inhibition of Wnt/β-Catenin Signaling by Novel Benzimidazole Compounds in Triple-Negative Breast Cancer. Int J Mol Sci 2018; 19. pii: E1524.

Schweizer MT, Haugk K, McKiernan JS, Gulati R, Cheng HH, Maes JL, et al. A phase I study of niclosamide in combination with enzalutamide in men with castration-resistant prostate cancer. PLoS One 2018; 13:e0198389.

Li R, Hu Z, Sun SY, Chen ZG, Owonikoko TK, Sica GL, et al. Niclosamide overcomes acquired resistance to erlotinib through suppression of STAT3 in non–small cell lung cancer. Mol Cancer Ther 2013; 12):2200-12.

Yo YT, Lin YW, Wang YC, Balch C, Huang RL, Chan MWY, et al. Growth inhibition of ovarian tumor–initiating cells by niclosamide. Mol Cancer Ther 2012; 11:1703-12.

Jin Y, Lu Z, Ding K, Li J, Du X, Chen C, et al. Antineoplastic mechanisms of niclosamide in acute myelogenous leukemia stem cells: inactivation of the NF-κB pathway and generation of reactive oxygen species. Cancer Res 2010; 70:2516-27.

Suliman MA, Zhang Z, Na H, Ribeiro AL, Zhang Y, Niang B, et al. Niclosamide inhibits colon cancer progression through downregulation of the Notch pathway and upregulation of the tumor suppressor miR-200 family. Int J Mol Med 2016; 38:776-84.

Chen L, Wang L, Shen H, Lin H, Li D. Anthelminthic drug niclosamide sensitizes the responsiveness of cervical cancer cells to paclitaxel via oxidative stress-mediated mTOR inhibition. Biochem Biophys Res Commun 2017; 484:416-21.

Chen M, Wang J, Lu J, Bond MC, Ren XR, Lyerly HK, et al. The anti-helminthic niclosamide inhibits Wnt/Frizzled1 signaling. Biochemistry 2009; 48:10267-74.

Osada T, Chen M, Yang XY, Spasojevic I, Vandeusen JB, Hsu D, et al. Antihelminth compound niclosamide downregulates Wnt signaling and elicits antitumor responses in tumors with activating APC mutations. Cancer Res 2011; 71:4172-82.

Kikuchi A, Yamamoto H, Kishida S. Multiplicity of the interactions of Wnt proteins and their receptors. Cell Signal 2007; 19:659-71.

MacDonald BT, Tamai K, He X. Wnt/β-catenin signaling: components, mechanisms, and diseases. Dev Cell 2009; 17:9-26.

Arend RC, Londoño-Joshi AI, Straughn JM, Buchsbaum DJ. The Wnt/β-catenin pathway in ovarian cancer: a review. Gynecol Oncol 2013; 131:772-9.

Barbolina MV, Burkhalter RJ, Stack MS. Diverse mechanisms for activation of Wnt signalling in the ovarian tumour microenvironment. Biochem J 2011; 437:1-12.

Boyer A, Goff AK, Boerboom D. WNT signaling in ovarian follicle biology and tumorigenesis. TEM 2010; 21:25-32.

Dakeng S, Duangmano S, Jiratchariyakul W, Bögler O, Patmasiriwat P. Inhibition of Wnt signaling by cucurbitacin B in breast cancer cells: Reduction of Wnt-associated proteins and reduced translocation of galectin‐3‐mediated β‐catenin to the nucleus. J Cell Biochem 2012; 113:49-60.

Anastas JN, Moon RT. WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer 2013; 13:11-26.

Mosmann, T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays methods. J Immunol Methods 1983; 16:55-63.

Organization WH. WHO Specifications and Evaluations for Public Health Pesticides (niclosamide). Geneva: World Health Organization; 2002; 1–24.

Chang YW, Yeh TK, Lin KT, Chen WC, Yao HT, Lan SJ, et al. Pharmacokinetics of anti-SARS-CoV agent niclosamide and Its analogs in rats. J Food Drug Anal 2006; 14:329–33.

Khanim FL, Merrick BAME, Giles HV, Jankute M, Jackson JB, Giles LJ, et al. Redeployment-based drug screening identifies the anti-helminthic niclosamide as anti-myeloma therapy that also reduces free light chain production. Blood Cancer J 2011; 1:e39.

Arend RC, Londoño-Joshi AI, Samant RS, Li Y, Conner M, Hidalgo B, et al. Inhibition of Wnt/β-catenin pathway by niclosamide: A therapeutic target for ovarian cancer. Gynecol Oncol 2014); 134:112-20.

Walters Haygood CL, Arend RC, Gangrade A, Chettiar S, Regan N, Hassmann CJ, et al. Niclosamide Analogs for Treatment of Ovarian Cancer. Int J Gynecol Cancer 2015; 25:1377-85.

King ML, Lindberg ME, Stodden GR, Okuda H, Ebers SD, Johnson A, et al. WNT7A/β-catenin signaling induces FGF1 and influences sensitivity to niclosamide in ovarian cancer. Oncogene 2015; 34:3452-62.

Published
2019-03-25
Section
Original Research