Nature Reviews Clinical Oncology. In cultured mammalian cells, stress conditions activate OMA1, leading to cleavage of L-OPA1 to S-OPA1 and inhibition of mitochondrial fusion, followed by mitochondrial division [23]. 2020; 11(12):3454-3466. F.G.S., Q.Z.C. Bethesda, MD 20894, Copyright Oral niclosamide inhibited tumor growth and progression in an intraperitoneal xenograft mouse model representative of human ovarian cancer. Niclosamide effectively suppresses ovarian carcinoma cell growth. 2011;71(12):4172-4182, 8. These data confirmed that Niclosamide has promising tumor-suppressive activity in ovarian carcinoma cells. In recent years, the survival of patients with ovarian cancer has been prolonged by the adoption of targeted therapy for poly ADP-ribose polymerase (PARP) vascular endothelial growth factor receptor (VEGFR) [2, 3]. Liu J, Nicum S, Reichardt P. et al. Briefly, SKOV3 and HO8910 cells were washed with ice-cold PBS and lysed in RIPA lysis buffer supplemented with protease and phosphatase inhibitors on ice for 20 min, followed by centrifugation at 13,000 rpm for 30 min at 4°C, and the supernatants were collected. Aerobic glycolysis alterations in response to nNiclosamide. These results suggest that using niclosamide is an attractive strategy to suppress the growth of ovarian cancer cells with aberrant Wnt signaling activation. Colonies were counted by two independent investigators. We also found that the cell growth inhibition in Niclosamide-treated, glucose-deprived cells was also rescued by NAC (10 mM) administration (Figure 4E). F. Western blot analysis of p-JNK and JNK in Niclosamide treated SKOV3 cells with or without NAC and in the presence or absence of glucose for 12 hr. CA: A Cancer Journal for Clinicians. Data are presented as mean ± SD. Received 2019-10-22 Accepted 2020-2-26 Published 2020-3-13, ©2021 Ivyspring International Publisher. Alexiou A, Soursou G, Chatzichronis S. et al. com; Prof. Bin Lu, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, University-Town, Wenzhou, Zhejiang 325035, China; E-mail address: lubmito thismailbody4 = thismailbody4 + "4;"; 2011;1(10):e39, 14. Flow cytometry analysis of cell apoptosis after the ovarian carcinoma cells treated with Niclosamide in the presence or absence of glucose for 12 hr. Ther Adv Med Oncol. The final concentrations of Oligomycin, FCCP, and rotenone were 0.1 μM. Interestingly, the differences in apoptosis induced by treatment with Niclosamide (8 μM) alone or by glucose deprivation alone were less than 10%. The Wnt/β-catenin pathway is known to regulate cellular proliferation and plays a role in chemoresistance. Therefore, we investigated the cytotoxicity of niclosamide analogs in combination with carboplatin against ovarian cancer patient ascites cells and tissue slices. 2019;129(8):3006-3017, 22. Would you like email updates of new search results? B. The next day, both SKOV3 and HO8910 cells were treated with gradient concentrations of Niclosamide (0, 4, and 8 μM) for 8 hr. Niclosamide, an old antihelminthic agent, demonstrates antitumor activity by blocking multiple signaling pathways of cancer stem cells. Niclosamide, an FDA approved salicyclamide derivative used for the treatment of tapeworm infections, targets the Wnt/β-catenin pathway. Background: Ovarian carcinoma is a common malignant tumor of the female reproductive organs with an incidence rate second only to cervical and endometrial cancers. Therefore, we investigated the cytotoxicity of niclosamide analogs in combination with carboplatin against ovarian cancer patient ascites cells and tissue slices. C. Western blot analysis of p-JNK and JNK in Niclosamide treated SKOV3 cells in the presence or absence of glucose for 12 hr. This study demonstrates that niclosamide is a potent Wnt/β-catenin inhibitor. This work was supported by the National Natural Science Foundation of China (81672757), Project of Science and Technology of Yantai (2016ZH082). 2018;150(1):173-179, 4. All statistical analyses were performed with the SPSS 16.0 statistical software package (SPSS Standard version 16.0, SPSS Inc., Chicago, IL). analyzed the data. Niclosamide is an "old" antihelminthic drug that uncouples mitochondria of intestinal parasites. In the present studies, Niclosamide showed tumor-suppressive activity in SKOV3 and HO8910 ovarian cancer cells as confirmed by a dose-dependent decrease in cell viability (Figure 1A). Targeting the Wnt/β-catenin pathway led to decreased cellular proliferation and increased cell death. 2020 Apr;301(4):895-899. doi: 10.1007/s00404-020-05510-9. thismailbody4 = thismailbody4 + ""; 1928;68(1767):437-443, 20. Niclosamide treatment (2 μM) of SKOV3 cells cultured under glucose deprivation conditions resulted in notable cell apoptosis when compared with cells cultured with glucose in the presence or absence of Niclosamide (2 μM) (Figure 4A and B). D. Representative morphological changes of SKOV3 and HO8910 cells in response to different concentrations of Niclosamide. Targeting the Wnt/β-catenin pathway led to decreased cellular proliferation and increased cell death. Terms of use. Higuchi T, Flies D B, Marjon N A. et al. 2014;5(4):1281-1294, 26. Copyright © 2014 Elsevier Inc. All rights reserved. Jin Y, Lu Z, Ding K. et al. The cellular bioenergetics data suggested that glucose deprivation could enhance the anti-tumor activity of Niclosamide. Discovery of new therapeutic approaches for treatment of metastatic breast cancer is still needed. In acute myelogenous leukemia and primary human glioblastoma cells, Niclosamide suppresses TAK1→IKK→IκBα→NF-κB signal transduction cascades and the expression of cleaved activated Notch1 receptor [14, 15]. Cancer statistics. G. SKOV3 and HO8910 cells were treated with or without Niclosamide (4, 8μM) for 24 hr. Giemsa and crystal violet were purchased from Solarbio Bioscience & Technology (Shanghai, China). The Seahorse bioenergetics analysis data confirmed that Niclosamide dramatically inhibited the overall mitochondrial oxygen consumption rate (OCR) (Figure 2A). School of Medicine, University of California - San Diego, La Jolla, CA 92037, USA6. E. The mitochondrial morphology in Niclosamide treated SKOV3 and HO8910 cells were examined by DAPI and MitoTracker Red. The role of other MAPK molecules (such as the p38 MAPK) in this process needs further confirmation. Representative images of colonies and total colonies were recorded and calculated. 2018;1859(4):263-269, 24. For glycolytic metabolism measurements, cells were incubated in basal media prior to injections using the Glycolytic Test kit (Seahorse Bioscience). thismailbody3 = thismailbody1; A and B. Protein Quality Control and Diseases Laboratory, Attardi Institute of Mitochondrial Biomedicine, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, P.R. This indicates that Niclosamide is more effective at inducing cell apoptosis under glucose restriction and suggests that a novel strategy for ovarian carcinoma therapy might be a combination of Niclosamide and a glycolysis inhibitor (such as 2-DG). 2015;25(8):1377-1385, 18. Thus, niclosamide has been reported as an anti-cancer molecule in numerous cancer types like CRC [ 13 ], osteosarcoma [ 42 ], breast cancer [ 30, 37 ], ovarian cancer [ 31, 43 ], prostate cancer [ 44 ], non-small cell lung cancer (NSCLC) [ 45, 46 ], glioblastoma [ 40 ], head and neck cancer [ 47 ], multiple myeloma, and leukemia [ 41, 48 ]. Biochemistry. A functional mammalian target of rapamycin complex 1 signaling is indispensable for c-Myc-driven hepatocarcinogenesis. 2015 Oct;25(8):1377-85. doi: 10.1097/IGC.0000000000000506. The most common type of ovarian cancer is epithelial cancer, followed by malignant germ cell tumors. SKOV3 cells were cultured in RPMI-1640 medium (Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (FBS, Gemini, USA) and antibiotics (100 units/mL penicillin and 100 units/mL streptomycin) and HO8910 cells were cultured in DMEM medium supplemented with 10% fetal bovine serum and antibiotics. JNK (c-Jun N-terminal kinase), also known as stress-activated protein kinase (SAPK), is a crucial member of the mitogen-activated protein kinase (MAPK) family in mammalian cells. Cells were collected and stained with Annexin V-fluorescein isothiocyanate (FITC) and PI. The effects of Niclosamide on the proliferation of SKOV3 and HO8910 cell lines were evaluated using Cell Counting Kit-8 (CCK-8). Objective. Methods: CCK-8 colony formation assays were performed to evaluate cell viability and tumor growth. thismailbody3 = thismailbody3+"2e;"; thismailbody4 = thismailbody4 + "4;"; The absorbance at 450 nm was measured using a Varioskan Flash microplate reader (Thermo Scientific, Waltham, MA). Khanim F L, Merrick B A M E, Giles H V. et al. document.write(thismailbody4 + thismailbody3+thismailbody2) 2012 Apr;31(4):178-84. doi: 10.5732/cjc.011.10290. var thismailbody3 = ""; Mol Cell. BCA protein assay kit and Pierce ECL western blotting substrate were obtained from Beyotime Biotechnology (Shanghai, China). Several studies have evaluated the anti-tumor activity of Niclosamide in suppressing prostate cancer, breast cancer, osteosarcoma, and colorectal carcinoma. The intact cellular oxygen consumption rate (OCR) and the extracellular acidification rate (ECAR) in compound treated SKOV3 and HO8910 cells were measured using a Seahorse XF-96 Extracellular Flux Analyzer (Seahorse Bioscience, North Billerica, MA) as described previously. eCollection 2020. The Anti-Helminthic Niclosamide inhibits Wnt/Frizzled1 signaling. //--> Londono-Joshi A I, Arend R C, Aristizabal L. et al. 2014;13:800-11 17. Hepatology. The Wnt/β-catenin pathway in ovarian cancer: a review. 2014;349(1):8-14, 19. Cellular Signalling. This would then increase HIF1-α expression and ultimately enhance glycolysis to overcome the shortage of ATP and other metabolites. Recent Patents on Anti-Cancer Drug Discovery. C. SKOV3 and HO8910 cells were treated with different concentrations of Niclosamide and cultured for 3 days. We also investigated the effects of increased basal glycolysis on ovarian carcinoma cell survival. We also explored potential mechanisms for the in vivo suppression of tumor growth by Niclosamide by western blot analysis of tumor sections. var thismailbody12 = "wm"; Gynecol Oncol. Gynecologic Oncology. Actin was used as a loading control. niclosamide has been identified as a direct inhibitor of signal transducer and activator of transcription 3 (STAT3) through interaction with the DNA-binding domain (37). To further confirm the effect of ERK1/2 inhibition on cell growth, ERK1/2 specific inhibitor SCH772984 was used to treat SKOV3 and HO8910 cells, we found ERK1/2 was significantly inactivated and cell growth was decreased (Figure S1A and B). The finding that Niclosamide decreased the basal cellular oxygen consumption rate while increasing the basal glycolytic rate indicated a stress response of metabolic reprogramming. Tissue or cell extracts equivalent to 20 μg total protein were resolved in 10% SDS-PAGE gels followed by electrophoretic transfer onto PVDF membrane (0.22 μM, Bio-Rad, Hercules, CA) in Tris-glycine buffer. All animal studies were performed with a protocol approved by the Institutional Animal Care and Use Committee of Wenzhou Medical University. Finally, slides were examined with White Light Laser Confocal Microscope Leica TCS SP8 X (Leica) and the fluorescence images were collected using a Delta Vision OMX 3D-strucured illumi-nation microscope. We found that abolishment of the excess ROS production with NAC (10 mM) abrogated the Niclosamide-induced cell apoptosis under glucose deprivation (Figure 4D). Niclosamide promotes cell apoptosis effectivelly under the condition of glucose deprivation. Dimethyl sulfoxide (DMSO) was used as a vehicle control. Epub 2020 Jul 21. Purpose: Adrenocortical carcinoma (ACC) is a rare and aggressive cancer, and no current effective therapy is available for locally advanced and metastatic ACC. Antibodies against ERK1/2 (67170-1-Ig), K-RAS (12063-1-Ig) and OMA1 (17116-1-Ig) were purchased from Proteintech (Wuhan, China). DAPI (C1002) and NAC (616-91-1) were purchased from Beyotime (Shanghai, China). Cells were then treated with different concentrations (0, 0.5, 1, 2, 4, 8, 16, or 32 μM) of Niclosamide or DMSO (vehicle control) for 48 hr, followed by Niclosamide incubating with CCK-8 for another 3 hr at 37°C. Prednisone is a drug that can help lessen inflammation. Combination treatment produced increased cytotoxicity compared to single agent treatment in 32/34 patient samples. and S.D.Y., wrote the paper. Dancey Janet. Each assay was performed in triplicate and data was derived from at least three independent experiments. Targeting metabolic reprograming can therefore significantly inhibit tumor growth and promote apoptosis. Cells were then treated with gradient concentrations (0, 4, and 8 μM) of Niclosamide for 1, 2, 3 and 4 days, and followed by incubating with Cell Counting Kit-8 (CCK-8) for 3 hr at 37°C, respectively. Phase Ib Study of Wnt Inhibitor Ipafricept with Gemcitabine and nab-paclitaxel in Patients with Previously Untreated Stage IV Pancreatic Cancer. A. Cells were collected and stained with Annexin V-fluorescein isothiocyanate (FITC) and PI. Giving abiraterone acetate, niclosamide, and prednisone may be a better treatment for patients with castration-resistant prostate cancer. High-throughput screening data showed that Niclosamide is an inhibitor of Wnt/β-catenin signaling and disrupts β-catenin/TCF complex formation while promoting Wnt co-receptor LRP6 degradation [6-9]. Redeployment-based drug screening identifies the anti-helminthic Niclosamide as anti-myeloma therapy that also reduces free light chain production. The basal glycolysis was increased in the Niclosamide-treated cells, whereas the glycolytic capacity and glycolytic reverse were both decreased (Figure 3A-D). Data are presented as mean ± SD. Niclosamide is an inexpensive and safe FDA-approved oral chlorinated salicylanilide antihelminthic/teniacidal agent with potential anticancer activity suggested in … Niclosamide also initiated apoptosis in a pool of SKOV3 and HO8910 cells, suggesting a further mechanism to explain Niclosamide suppression of cancer cell growth (Figure 1F). Protease (Complete Mini) and phosphatase (PhosphoSTOP) inhibitor cocktail tablets were purchased from Roche Applied Science (Indianapolis, IN). The data expressed in right graphs represent the mean ± SD. Results. Tumor dimensions were determined using calipers, and the tumor volume (mm3) was calculated using the following the formula: volume = length × (width) 2/2. FOIA ALDH1A1 and LRP6 were analyzed by IHC in solid tumor and in ascites before and after treatment with niclosamide. D. Flow cytometry analysis of cell apoptosis after the ovarian carcinoma cells were treated with Niclosamide and/or NAC in the presence or absence of glucose for 12 hr. Niclosamide, a salicylamide derivative, was originally an anti-helminthic drug but more recently has been explored in variety of malignancies including HCC, colon cancer, and ovarian cancer 4-7. 2017;6(2):174-184, 27. International Journal of Gynecological Cancer. Effect of niclosamide on basal-like breast cancers. B. SKOV3 cells were treated with Niclosamide in the presence or absence of glucose for 12 hr, then subjected to culture for 3 days. Niclosamide, a drug used to deal with tapeworm infections, attacks cells with a p53 deficiency, a deficiency that the majority of cancer cells possess, causing apoptosis or cancer cell death and reducing tumour size by up to 50%; it can also attack restrict metastasis and cancer cell migration and inhibit cancer stem cells. Data are presented as mean ± SE (n = 6). var thismailbody11 = ""; A Seahorse XF96 analyzer was used to measure cellular bioenergetics. However, an unanswered question is whether Niclosamide has a role in the regulation of other signal pathways with known involvement in cancer progression. China4. Role of GTPases in the Regulation of Mitochondrial Dynamics in Alzheimer's Disease and CNS-Related Disorders. Specifically, L-OPA1, but not S-OPA1, is essential for mitochondrial fusion, whereas accumulation of S-OPA1 promotes mitochondrial division. A previous study showed that c-Myc up-regulates SLC1A5 to promote amino acid metabolism, which subsequently activates mTORC1 signaling to favor hepatocarcinogenesis [24, 25]. Female athymic nude mice were purchased from Shanghai Laboratory Animal Center, CAS (Shanghai, China) and housed in a specific pathogen-free (SPF) environment. PLoS One. Briefly, SKOV3 andHO8910 cells were seeded into 96-well plates at a density of 3×103 cells/well and incubated overnight at 37°C with 5% CO2. Objective Niclosamide has shown activity against ovarian cancer in vitro; however, it has low bioavailability in vivo. The primary antibodies against HK2 (500994), PFKM (505477), PGK1 (501965), and LDHA (501146) were obtained from Zen Bioscience (Chengdu, China). Actin was used as a loading control. Cao Y. Adipocyte and lipid metabolism in cancer drug resistance. Niclosamide (amuricide or molluscicide) is an anthelmintic salicylamide derivative that inhibits mitochondrial oxidative phosphorylation in the cells of parasitic worms, thereby restricting ATP production. E. Changes in body weight in saline- or Niclosamide-treated mice. Data are presented as mean ± SD. For apoptosis analysis, the SKOV3 and HO8910 cells were treated with gradient concentrations of Niclosamide (0, 4, and 8 μM) for 24 hr or with Niclosamide and/or NAC (10mM) in the presence or absence of glucose for 12 hr; cells were then collected and incubated with Annexin V-FITC/ PI (BD, San Jose, CA) in the dark at room temperature for 20 min, according to the manufacturer's protocol. 2015;3(11):1257-1268, 5. The extracellular acidification rate (ECAR) of Niclosamide treated SKOV3 and HO8910 cells under the indicated conditions were measured in real time using the Seahorse XF96 Extracellular Flux Analyzer. Dotan E, Cardin DB, Lenz HJ, Messersmith W, O'Neil B, Cohen SJ, Denlinger CS, Shahda S, Astsaturov I, Kapoun AM, Brachmann RK, Uttamsingh S, Stagg RJ, Weekes C. Clin Cancer Res. Mitotracker (#9082) was purchased from Cell Signaling Technology (Beverly, MA). Key Laboratory of Diagnosis and Treatment of Severe Hepato-Pancreatic Diseases of Zhejiang Province, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China.3. Keywords: Niclosamide, ovarian carcinoma, cellular bioenergetics, MEK1/2-ERK1/2 signal. Our data using the ROS scavenger NAC also confirmed that Niclosamide-induced apoptosis under glucose deprivation is ROS-dependent. Keywords: 2014;13(4):800-811, 10. Cancer stem cells; Chemoresistance; LRP6; Niclosamide; Ovarian cancer; Wnt/β-catenin. Int J Gynecol Cancer. Dr. Otto Warburg found tumor cells dependent on glycolysis to support cell growth even under normoxic condition which named “Warburg effect”. Niclosamide significantly represses mitochondrial respiration and interrupt mitochondrial dynamics. Corresponding author: Prof. Qizhi Cao, School of Basic Medical Sciences, Binzhou Medical University, No. Both SKOV3 and HO8910 cells were counted and a total of 800 cells per well were seeded evenly into 6-well plates and incubated at 37°C for 7-10 days in a humidified incubator with 5% CO2. In patients with ovarian epithelial cancer, less than 30% of the tumors are confined to the ovary, and most of tumors spread to the pelvic and abdominal organs, thereby further complicating early diagnosis. Data are presented as mean ± SE (n = 6). Collectively, these results indicate that FGF1 is a direct downstream target of WNT7A/β-catenin signaling and this pathway has potential as a therapeutic target in ovarian cancer. 2020 May 26;13(1):60. doi: 10.1186/s13045-020-00901-6. Mol Cancer Ther. Biochimica Biophysica Acta (BBA)-Bioenergetics. Tumorspheres were treated, and Wnt transcriptional activity was measured by the TOPflash reporter assay. Objective Niclosamide has shown activity against ovarian cancer in vitro; however, it has low bioavailability in vivo. To further confirm the effect of Niclosamide on OPA1 cleavage in vivo, we analyzed OPA1 level in mock and Niclosamide administrated nude mice tumor sections and found OPA1 was cleaved which was consistent with the data in vitro (Figure S5A). Glucose metabolism, glutamine metabolism, and lipid metabolism are the most significant metabolic pathways that commonly reprograming in tumor cells.