Skip to main content
Top
Published in: Tumor Biology 3/2016

01-03-2016 | Original Article

Apoptotic effects of salinomycin on human ovarian cancer cell line (OVCAR-3)

Authors: Fuat Kaplan, Fulya Teksen

Published in: Tumor Biology | Issue 3/2016

Login to get access

Abstract

In this study, we studied the apoptotic and cytotoxic effects of salinomycin on human ovarian cancer cell line (OVCAR-3) as salinomycin is known as a selectively cancer stem cell killer agent. We used immortal human ovarian epithelial cell line (IHOEC) as control group. Ovarian cancer cells and ovarian epithelial cells were treated by different concentrations of salinomycin such as 0.1, 1, and 40 μM and incubated for 24, 48, and 72 h. Dimethylthiazol (MTT) cell viability assay was performed to determine cell viability and toxicity. On the other hand, the expression levels of some of the apoptosis-related genes, namely anti-apoptotic Bcl-2, apoptotic Bax, and Caspase-3 were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Additionally, Caspase-3 protein level was also determined. As a result, we concluded that incubation of human OVCAR-3 by 0.1 μM concentration of salinomycin for 24 h killed 40 % of the cancer cells by activating apoptosis but had no effect on normal cells. The apoptotic Bax gene expression was upregulated but anti-apoptotic Bcl-2 gene expression was downregulated. Active Caspase-3 protein level was increased significantly (p < 0.05).
Literature
2.
go back to reference Aletti GD et al. Current management strategies for ovarian cancer. Mayo Clin Proc. 2007;82(6):751–70.CrossRefPubMed Aletti GD et al. Current management strategies for ovarian cancer. Mayo Clin Proc. 2007;82(6):751–70.CrossRefPubMed
3.
go back to reference Eum KH, Lee M. Crosstalk between autophagy and apoptosis in the regulation of paclitaxel-induced cell death in v-Ha-ras-transformed fibroblasts. Mol Cell Biochem. 2011;348(1–2):61–8.CrossRefPubMed Eum KH, Lee M. Crosstalk between autophagy and apoptosis in the regulation of paclitaxel-induced cell death in v-Ha-ras-transformed fibroblasts. Mol Cell Biochem. 2011;348(1–2):61–8.CrossRefPubMed
4.
go back to reference Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972;26(4):239–57.CrossRefPubMedPubMedCentral Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972;26(4):239–57.CrossRefPubMedPubMedCentral
5.
go back to reference Engel T, Henshall DC. Apoptosis, Bcl-2 family proteins and caspases: the ABCs of seizure-damage and epileptogenesis? Int J Physiol Pathophysiol Pharmacol. 2009;1(2):97–115.PubMedPubMedCentral Engel T, Henshall DC. Apoptosis, Bcl-2 family proteins and caspases: the ABCs of seizure-damage and epileptogenesis? Int J Physiol Pathophysiol Pharmacol. 2009;1(2):97–115.PubMedPubMedCentral
6.
go back to reference Ghobrial IM, Witzig TE, Adjei AA. Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin. 2005;55(3):178–94.CrossRefPubMed Ghobrial IM, Witzig TE, Adjei AA. Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin. 2005;55(3):178–94.CrossRefPubMed
7.
go back to reference Tsujimoto Y. Role of Bcl-2 family proteins in apoptosis: apoptosomes or mitochondria? Genes Cells. 1998;3(11):697–707.CrossRefPubMed Tsujimoto Y. Role of Bcl-2 family proteins in apoptosis: apoptosomes or mitochondria? Genes Cells. 1998;3(11):697–707.CrossRefPubMed
8.
9.
go back to reference Mitani M, Yamanishi T, Miyazaki Y. Salinomycin: a new monovalent cation ionophore. Biochem Biophys Res Commun. 1975;66(4):1231–6.CrossRefPubMed Mitani M, Yamanishi T, Miyazaki Y. Salinomycin: a new monovalent cation ionophore. Biochem Biophys Res Commun. 1975;66(4):1231–6.CrossRefPubMed
10.
go back to reference Butaye P, Devriese LA, Haesebrouck F. Antimicrobial growth promoters used in animal feed: effects of less well known antibiotics on gram-positive bacteria. Clin Microbiol Rev. 2003;16(2):175–88.CrossRefPubMedPubMedCentral Butaye P, Devriese LA, Haesebrouck F. Antimicrobial growth promoters used in animal feed: effects of less well known antibiotics on gram-positive bacteria. Clin Microbiol Rev. 2003;16(2):175–88.CrossRefPubMedPubMedCentral
11.
go back to reference Callaway TR et al. Ionophores: their use as ruminant growth promotants and impact on food safety. Curr Issues Intest Microbiol. 2003;4(2):43–51.PubMed Callaway TR et al. Ionophores: their use as ruminant growth promotants and impact on food safety. Curr Issues Intest Microbiol. 2003;4(2):43–51.PubMed
12.
go back to reference Danforth HD et al. Anticoccidial activity of salinomycin in battery raised broiler chickens. Poult Sci. 1977;56(3):926–32.CrossRefPubMed Danforth HD et al. Anticoccidial activity of salinomycin in battery raised broiler chickens. Poult Sci. 1977;56(3):926–32.CrossRefPubMed
14.
go back to reference Bortner CD, Hughes Jr FM, Cidlowski JA. A primary role for K+ and Na+ efflux in the activation of apoptosis. J Biol Chem. 1997;272(51):32436–42.CrossRefPubMed Bortner CD, Hughes Jr FM, Cidlowski JA. A primary role for K+ and Na+ efflux in the activation of apoptosis. J Biol Chem. 1997;272(51):32436–42.CrossRefPubMed
15.
go back to reference Fuchs D et al. Salinomycin overcomes ABC transporter-mediated multidrug and apoptosis resistance in human leukemia stem cell-like KG-1a cells. Biochem Biophys Res Commun. 2010;394(4):1098–104.CrossRefPubMed Fuchs D et al. Salinomycin overcomes ABC transporter-mediated multidrug and apoptosis resistance in human leukemia stem cell-like KG-1a cells. Biochem Biophys Res Commun. 2010;394(4):1098–104.CrossRefPubMed
16.
go back to reference Fuchs D et al. Salinomycin induces apoptosis and overcomes apoptosis resistance in human cancer cells. Biochem Biophys Res Commun. 2009;390(3):743–9.CrossRefPubMed Fuchs D et al. Salinomycin induces apoptosis and overcomes apoptosis resistance in human cancer cells. Biochem Biophys Res Commun. 2009;390(3):743–9.CrossRefPubMed
17.
go back to reference Morgan DM. Tetrazolium (MTT) assay for cellular viability and activity. Methods Mol Biol. 1998;79:179–83.PubMed Morgan DM. Tetrazolium (MTT) assay for cellular viability and activity. Methods Mol Biol. 1998;79:179–83.PubMed
18.
go back to reference Abu-Qare AW, Abou-Donia MB. Biomarkers of apoptosis: release of cytochrome c, activation of caspase-3, induction of 8-hydroxy-2′-deoxyguanosine, increased 3-nitrotyrosine, and alteration of p53 gene. J Toxicol Environ Health B Crit Rev. 2001;4(3):313–32.CrossRefPubMed Abu-Qare AW, Abou-Donia MB. Biomarkers of apoptosis: release of cytochrome c, activation of caspase-3, induction of 8-hydroxy-2′-deoxyguanosine, increased 3-nitrotyrosine, and alteration of p53 gene. J Toxicol Environ Health B Crit Rev. 2001;4(3):313–32.CrossRefPubMed
19.
go back to reference Park IS, Kim JE. Potassium efflux during apoptosis. J Biochem Mol Biol. 2002;35(1):41–6.PubMed Park IS, Kim JE. Potassium efflux during apoptosis. J Biochem Mol Biol. 2002;35(1):41–6.PubMed
20.
go back to reference Parajuli B et al. Salinomycin inhibits Akt/NF-kappaB and induces apoptosis in cisplatin resistant ovarian cancer cells. Cancer Epidemiol. 2013;37(4):512–7.CrossRefPubMed Parajuli B et al. Salinomycin inhibits Akt/NF-kappaB and induces apoptosis in cisplatin resistant ovarian cancer cells. Cancer Epidemiol. 2013;37(4):512–7.CrossRefPubMed
21.
go back to reference Kim KY et al. Salinomycin-induced apoptosis of human prostate cancer cells due to accumulated reactive oxygen species and mitochondrial membrane depolarization. Biochem Biophys Res Commun. 2011;413(1):80–6.CrossRefPubMed Kim KY et al. Salinomycin-induced apoptosis of human prostate cancer cells due to accumulated reactive oxygen species and mitochondrial membrane depolarization. Biochem Biophys Res Commun. 2011;413(1):80–6.CrossRefPubMed
22.
23.
go back to reference Zhou J et al. Salinomycin induces apoptosis in cisplatin-resistant colorectal cancer cells by accumulation of reactive oxygen species. Toxicol Lett. 2013;222(2):139–45.CrossRefPubMed Zhou J et al. Salinomycin induces apoptosis in cisplatin-resistant colorectal cancer cells by accumulation of reactive oxygen species. Toxicol Lett. 2013;222(2):139–45.CrossRefPubMed
24.
go back to reference Xu S-q, Z A-z, Liu C-c, Xu T-t, Chen X-y, Liu G-x. Salinomycin inhibits proliferation and induces apoptosis of Gleevec-resistant chronic myeloid leukemic cell line K562/Glv. Chin J Pathophysiol. 2012;28:1208–1212. Xu S-q, Z A-z, Liu C-c, Xu T-t, Chen X-y, Liu G-x. Salinomycin inhibits proliferation and induces apoptosis of Gleevec-resistant chronic myeloid leukemic cell line K562/Glv. Chin J Pathophysiol. 2012;28:1208–1212.
25.
go back to reference Zeng J, Liu C-c, Zhu A-z, Chen X-y, Tan G-x, Liu G-x. Salinomycin inhibited proliferation and induced apoptosis of cisplatin-resistant human lung adenocarcinoma cell line A549/DDP.pdf. Chin J Pathophysiol. 2012;28:834–8. Zeng J, Liu C-c, Zhu A-z, Chen X-y, Tan G-x, Liu G-x. Salinomycin inhibited proliferation and induced apoptosis of cisplatin-resistant human lung adenocarcinoma cell line A549/DDP.pdf. Chin J Pathophysiol. 2012;28:834–8.
26.
go back to reference Wu D et al. Salinomycin inhibits proliferation and induces apoptosis of human nasopharyngeal carcinoma cell in vitro and suppresses tumor growth in vivo. Biochem Biophys Res Commun. 2014;443(2):712–7.CrossRefPubMed Wu D et al. Salinomycin inhibits proliferation and induces apoptosis of human nasopharyngeal carcinoma cell in vitro and suppresses tumor growth in vivo. Biochem Biophys Res Commun. 2014;443(2):712–7.CrossRefPubMed
27.
go back to reference Wang F et al. Salinomycin inhibits proliferation and induces apoptosis of human hepatocellular carcinoma cells in vitro and in vivo. PLoS One. 2012;7(12):e50638.CrossRefPubMedPubMedCentral Wang F et al. Salinomycin inhibits proliferation and induces apoptosis of human hepatocellular carcinoma cells in vitro and in vivo. PLoS One. 2012;7(12):e50638.CrossRefPubMedPubMedCentral
28.
go back to reference Shin S-J et al. Salinomycin have antiproliferative and apoptotic effects on ovarian cancer stem-like cell. In: Proceedings of the 105th annual meeting of the american association for cancer research. 2014. Cancer Res 2014: 5–9. Shin S-J et al. Salinomycin have antiproliferative and apoptotic effects on ovarian cancer stem-like cell. In: Proceedings of the 105th annual meeting of the american association for cancer research. 2014. Cancer Res 2014: 5–9.
Metadata
Title
Apoptotic effects of salinomycin on human ovarian cancer cell line (OVCAR-3)
Authors
Fuat Kaplan
Fulya Teksen
Publication date
01-03-2016
Publisher
Springer Netherlands
Published in
Tumor Biology / Issue 3/2016
Print ISSN: 1010-4283
Electronic ISSN: 1423-0380
DOI
https://doi.org/10.1007/s13277-015-4212-6

Other articles of this Issue 3/2016

Tumor Biology 3/2016 Go to the issue
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine