Anticancer Effects of Cold Atmospheric Plasma in Canine Osteosarcoma Cells
Abstract
:1. Introduction
2. Results
2.1. CAP Generation by Dielectric Barrier Discharge
2.2. CAP Induces Cell Growth Arrest in Canine Osteosarcoma Cell Lines
2.3. CAP Generates ROS and Causes DNA Damage in A Canine Osteosarcoma Cell Line
2.4. CAP Induces Apoptosis in a Canine Osteosarcoma Cell Line
2.5. CAP-Treated Canine Osteosarcoma Cells Showed Decreased Invasion and Migration Activity
3. Discussion
4. Materials and Methods
4.1. Cell Cultures and CAP Induction
4.2. ROS Detection
4.3. Western Blot Analysis
4.4. Cell Viability Assay
4.5. De novo DNA Synthesis Detection
4.6. Apoptotic Bodies and Mitochondrial Membrane Potential Detection
4.7. Detection of Apoptotic Cells Using Flow Cytometry
4.8. Cell Migration Assay
4.9. Three-Dimensional Tumor Spheroid Invasion Assay
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
CAP | Cold atmospheric plasma |
DBD | Dielectric-barrier discharge |
HCS | High-content screening |
ROS | Reactive oxygen species |
EdU | 5-ethynyl-2′-deoxyuridine |
H2DCFDA | 2′,7′-dichlorodihydrofluorescein diacetate |
DCF | 2′,7′-dichlorodihydrofluorescein |
DAPI | 4′,6-diamidino-2-phenylindole |
References
- Simpson, S.; Dunning, M.D.; De Brot, S.; Grau-Roma, L.; Mongan, N.P.; Rutland, C.S. Comparative review of human and canine osteosarcoma: Morphology, epidemiology, prognosis, treatment and genetics. Acta Vet. Scand. 2017, 59, 71. [Google Scholar] [CrossRef] [PubMed]
- Frimberger, A.; Chan, C.M.; Moore, A.S. Canine Osteosarcoma Treated by Post-Amputation Sequential Accelerated Doxorubicin and Carboplatin Chemotherapy: 38 Cases. J. Am. Anim. Hosp. Assoc. 2016, 52, 149–156. [Google Scholar] [CrossRef] [PubMed]
- Nolan, M.W.; Gieger, T. Update in Veterinary Radiation Oncology. Vet. Clin. N. Am. Small Anim. Pr. 2019, 49, 933–947. [Google Scholar] [CrossRef] [PubMed]
- Kubicek, L.; VanderHart, D.; Wirth, K.; An, Q.; Chang, M.; Farese, J.; Bova, F.; Sudhyadhom, A.; Kow, K.; Bacon, N.J.; et al. Association between computed tomographic characteristics and fractures following stereotactic radiosurgery in dogs with appendicular osteosarcoma. Vet. Radiol. Ultrasound 2016, 57, 321–330. [Google Scholar] [CrossRef]
- Kong, M.G.; Kroesen, G.; Morfill, G.; Nosenko, T.; Shimizu, T.; Van Dijk, J.; Zimmermann, J.L. Plasma medicine: An introductory review. New J. Phys. 2009, 11, 115012. [Google Scholar] [CrossRef]
- Yoon, Y.; Ku, B.; Lee, K.; Jung, Y.J.; Baek, S.J. Cold Atmospheric Plasma Induces HMGB1 Expression in Cancer Cells. Anticancer Res. 2019, 39, 2405–2413. [Google Scholar] [CrossRef]
- Braný, D.; Dvorská, D.; Halašová, E.; Škovierová, H. Cold Atmospheric Plasma: A Powerful Tool for Modern Medicine. Int. J. Mol. Sci. 2020, 21, 2932. [Google Scholar] [CrossRef] [Green Version]
- Bekeschus, S.; Clemen, R.; Metelmann, H.-R. Potentiating anti-tumor immunity with physical plasma. Clin. Plasma Med. 2018, 12, 17–22. [Google Scholar] [CrossRef]
- Semmler, M.L.; Bekeschus, S.; Schäfer, M.; Bernhardt, T.; Fischer, T.; Witzke, K.; Seebauer, C.; Rebl, H.; Grambow, E.; Vollmar, B.; et al. Molecular Mechanisms of the Efficacy of Cold Atmospheric Pressure Plasma (CAP) in Cancer Treatment. Cancers 2020, 12, 269. [Google Scholar] [CrossRef] [Green Version]
- Izadjoo, M.; Zack, S.; Kim, H.; Skiba, J. Medical applications of cold atmospheric plasma: State of the science. J. Wound Care 2018, 27, S4–S10. [Google Scholar] [CrossRef]
- Hirst, A.M.; Frame, F.M.; Arya, M.; Maitland, N.J.; O’Connell, D. Low temperature plasmas as emerging cancer therapeutics: The state of play and thoughts for the future. Tumor Biol. 2016, 37, 7021–7031. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kim, C.-H.; Bahn, J.H.; Lee, S.-H.; Kim, G.-Y.; Jun, S.-I.; Lee, K.; Baek, S.J. Induction of cell growth arrest by atmospheric non-thermal plasma in colorectal cancer cells. J. Biotechnol. 2010, 150, 530–538. [Google Scholar] [CrossRef] [PubMed]
- Kim, C.-H.; Kwon, S.; Bahn, J.H.; Lee, K.; Jun, S.I.; Rack, P.D.; Baek, S.J. Effects of atmospheric nonthermal plasma on invasion of colorectal cancer cells. Appl. Phys. Lett. 2010, 96, 243701. [Google Scholar] [CrossRef] [Green Version]
- Lin, A.G.; Xiang, B.; Merlino, D.J.; Baybutt, T.R.; Sahu, J.; Fridman, A.; Snook, A.E.; Miller, V. Non-thermal plasma induces immunogenic cell death in vivo in murine CT26 colorectal tumors. Oncoimmunology 2018, 7, e1484978. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Azzariti, A.; Iacobazzi, R.M.; Di Fonte, R.; Porcelli, L.; Gristina, R.; Favia, P.; Fracassi, F.; Trizio, I.; Silvestris, N.; Guida, G.; et al. Plasma-activated medium triggers cell death and the presentation of immune activating danger signals in melanoma and pancreatic cancer cells. Sci. Rep. 2019, 9, 4099. [Google Scholar] [CrossRef] [PubMed]
- Mitra, S.; Nguyen, L.N.; Akter, M.; Park, G.; Kaushik, N.; Kaushik, N. Impact of ROS Generated by Chemical, Physical, and Plasma Techniques on Cancer Attenuation. Cancers 2019, 11, 1030. [Google Scholar] [CrossRef] [Green Version]
- Bekeschus, S.; Lippert, M.; Diepold, K.; Chiosis, G.; Seufferlein, T.; Azoitei, N. Physical plasma-triggered ROS induces tumor cell death upon cleavage of HSP90 chaperone. Sci. Rep. 2019, 9, 4112. [Google Scholar] [CrossRef] [Green Version]
- Conway, G.E.; He, Z.; Hutanu, A.L.; Cribaro, G.P.; Manaloto, E.; Casey, A.; Traynor, D.; Milosavljevic, V.; Howe, O.; Barcia, C.; et al. Cold Atmospheric Plasma induces accumulation of lysosomes and caspase-independent cell death in U373MG glioblastoma multiforme cells. Sci. Rep. 2019, 9, 1–12. [Google Scholar] [CrossRef] [Green Version]
- Norbury, C.J.; Zhivotovsky, B. DNA damage-induced apoptosis. Oncogene 2004, 23, 2797–2808. [Google Scholar] [CrossRef] [Green Version]
- Hirst, A.M.; Simms, M.S.; Mann, V.M.; Maitland, N.J.; O’Connell, D.; Frame, F.M. Low-temperature plasma treatment induces DNA damage leading to necrotic cell death in primary prostate epithelial cells. Br. J. Cancer 2015, 112, 1536–1545. [Google Scholar] [CrossRef] [Green Version]
- Luetke, A.; Meyers, P.A.; Lewis, I.; Juergens, H. Osteosarcoma treatment—Where do we stand? A state of the art review. Cancer Treat. Rev. 2014, 40, 523–532. [Google Scholar] [CrossRef] [PubMed]
- Ohba, T.; Cates, J.M.M.; Cole, H.A.; Slosky, D.A.; Haro, H.; Ichikawa, J.; Ando, T.; Schwartz, H.S.; Schoenecker, J.G. Pleiotropic effects of bisphosphonates on osteosarcoma. Bone 2014, 63, 110–120. [Google Scholar] [CrossRef] [PubMed]
- Wycislo, K.; Fan, T.M. The Immunotherapy of Canine Osteosarcoma: A Historical and Systematic Review. J. Vet. Intern. Med. 2015, 29, 759–769. [Google Scholar] [CrossRef] [PubMed]
- Haralambiev, L.; Wien, L.; Gelbrich, N.; Bakir, S.; Kramer, A.; Burchardt, M.; Ekkernkamp, A.; Stope, M.B.; Lange, J.; Gümbel, D. Cold atmospheric plasma inhibits the growth of osteosarcoma cells by inducing apoptosis, independent of the device used. Oncol. Lett. 2019, 19, 283–290. [Google Scholar] [CrossRef] [Green Version]
- Gümbel, D.; Bekeschus, S.; Gelbrich, N.; Napp, M.; Ekkernkamp, A.; Kramer, A.; Stope, M.B. Cold Atmospheric Plasma in the Treatment of Osteosarcoma. Int. J. Mol. Sci. 2017, 18, 2004. [Google Scholar] [CrossRef] [Green Version]
- Liu, J.-R.; Xu, G.-M.; Shi, X.; Zhang, G. Low temperature plasma promoting fibroblast proliferation by activating the NF-κB pathway and increasing cyclinD1 expression. Sci. Rep. 2017, 7, 11698. [Google Scholar] [CrossRef] [Green Version]
- Tafani, M.; Sansone, L.; Limana, F.; Arcangeli, T.; De Santis, E.; Polese, M.; Fini, M.; Russo, M.A. The Interplay of Reactive Oxygen Species, Hypoxia, Inflammation, and Sirtuins in Cancer Initiation and Progression. Oxid. Med. Cell. Longev. 2016, 2016, 1–18. [Google Scholar] [CrossRef] [Green Version]
- Schito, L. Hypoxia-Dependent Angiogenesis and Lymphangiogenesis in Cancer. In Hypoxia and Cancer Metastasis; Gilkes, D.M., Ed.; Springer International Publishing: Cham, Switzerland, 2019; pp. 71–85. [Google Scholar]
- Galadari, S.; Rahman, A.; Pallichankandy, S.; Thayyullathil, F. Reactive oxygen species and cancer paradox: To promote or to suppress? Free Radic. Biol. Med. 2017, 104, 144–164. [Google Scholar] [CrossRef]
- Khan, S.A.; Smith, N.L.; Wilson, A.L.; Gandhi, J.; Vatsia, S. Ozone therapy: An overview of pharmacodynamics, current research, and clinical utility. Med Gas Res. 2017, 7, 212–219. [Google Scholar] [CrossRef] [Green Version]
- Simkó, M.; Mattsson, M.-O. Extremely low frequency electromagnetic fields as effectors of cellular responses in vitro: Possible immune cell activation. J. Cell. Biochem. 2004, 93, 83–92. [Google Scholar] [CrossRef]
- Chen, Z.; Simonyan, H.; Cheng, X.; Gjika, E.; Lin, L.; Canady, J.; Sherman, J.H.; Young, C.N.; Keidar, M. A Novel Micro Cold Atmospheric Plasma Device for Glioblastoma both In Vitro and In Vivo. Cancers 2017, 9, 61. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xiang, L.; Xu, X.; Zhang, S.; Cai, D.; Dai, X. Cold atmospheric plasma conveys selectivity on triple negative breast cancer cells both in vitro and in vivo. Free Radic. Biol. Med. 2018, 124, 205–213. [Google Scholar] [CrossRef] [PubMed]
- Szewczyk, M.; Lechowski, R.; Zabielska, K. What do we know about canine osteosarcoma treatment? Review. Vet. Res. Commun. 2014, 39, 61–67. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Park, S.; Kim, H.; Ji, H.W.; Kim, H.W.; Yun, S.H.; Choi, E.H.; Kim, S.J. Cold Atmospheric Plasma Restores Paclitaxel Sensitivity to Paclitaxel-Resistant Breast Cancer Cells by Reversing Expression of Resistance-Related Genes. Cancers 2019, 11, 2011. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lee, S.; Lee, H.; Jeong, D.; Ham, J.; Park, S.; Choi, E.H.; Kim, S.J. Cold atmospheric plasma restores tamoxifen sensitivity in resistant MCF-7 breast cancer cell. Free Radic. Biol. Med. 2017, 110, 280–290. [Google Scholar] [CrossRef]
- Köritzer, J.; Boxhammer, V.; Schafer, A.; Shimizu, T.; Klämpfl, T.G.; Li, Y.-F.; Welz, C.; Schwenk-Zieger, S.; Morfill, G.E.; Zimmermann, J.L.; et al. Restoration of Sensitivity in Chemo—Resistant Glioma Cells by Cold Atmospheric Plasma. PLoS ONE 2013, 8, e64498. [Google Scholar] [CrossRef] [Green Version]
- Argentiero, A.; Solimando, A.G.; Brunetti, O.; Calabrese, A.; Pantano, F.; Iuliani, M.; Santini, D.; Silvestris, N.; Vacca, A. Skeletal Metastases of Unknown Primary: Biological Landscape and Clinical Overview. Cancers 2019, 11, 1270. [Google Scholar] [CrossRef] [Green Version]
- Di Marzo, L.; DeSantis, V.; Solimando, A.G.; Ruggieri, S.; Annese, T.; Nico, B.; Fumarulo, R.; Vacca, A.; Frassanito, M.A. Microenvironment drug resistance in multiple myeloma: Emerging new players. Oncotarget 2016, 7, 60698–60711. [Google Scholar] [CrossRef] [Green Version]
- Khalili, M.; Daniels, L.; Lin, A.; Krebs, F.C.; Snook, A.E.; Bekeschus, S.; Bowne, W.B.; Miller, V. Non-thermal plasma-induced immunogenic cell death in cancer. J. Phys. D Appl. Phys. 2019, 52, 423001. [Google Scholar] [CrossRef]
- Lin, A.; Truong, B.; Patel, S.; Kaushik, N.K.; Kaushik, N.; Fridman, G.; Fridman, A.; Miller, V. Nanosecond-Pulsed DBD Plasma-Generated Reactive Oxygen Species Trigger Immunogenic Cell Death in A549 Lung Carcinoma Cells through Intracellular Oxidative Stress. Int. J. Mol. Sci. 2017, 18, 966. [Google Scholar] [CrossRef] [Green Version]
- Eggers, B.; Marciniak, J.; Memmert, S.; Kramer, F.J.; Deschner, J.; Nokhbehsaim, M. The beneficial effect of cold atmospheric plasma on parameters of molecules and cell function involved in wound healing in human osteoblast-like cells in vitro. Odontology 2020, 1–10. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Hartwig, S.; Doll, C.; Voss, J.O.; Hertel, M.; Preissner, S.; Raguse, J.D. Treatment of Wound Healing Disorders of Radial Forearm Free Flap Donor Sites Using Cold Atmospheric Plasma: A Proof of Concept. J. Oral Maxillofac. Surg. 2017, 75, 429–435. [Google Scholar] [CrossRef] [PubMed]
- Haertel, B.; Von Woedtke, T.; Weltmann, K.-D.; Lindequist, U. Non-Thermal Atmospheric-Pressure Plasma Possible Application in Wound Healing. Biomol. Ther. 2014, 22, 477–490. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ryu, S.; Park, S.; Lim, W.; Song, G. Quercetin augments apoptosis of canine osteosarcoma cells by disrupting mitochondria membrane potential and regulating PKB and MAPK signal transduction. J. Cell. Biochem. 2019, 120, 17449–17458. [Google Scholar] [CrossRef] [PubMed]
- Park, H.; Park, S.; Bazer, F.W.; Lim, W.; Song, G. Myricetin treatment induces apoptosis in canine osteosarcoma cells by inducing DNA fragmentation, disrupting redox homeostasis, and mediating loss of mitochondrial membrane potential. J. Cell. Physiol. 2018, 233, 7457–7466. [Google Scholar] [CrossRef] [PubMed]
- Yamaguchi, K.; Whitlock, N.C.; Liggett, J.L.; Legendre, A.M.; Fry, M.; Baek, S.J. Molecular characterisation of canine nonsteroidal anti-inflammatory drug-activated gene (NAG-1). Vet. J. 2007, 175, 89–95. [Google Scholar] [CrossRef] [Green Version]
- Lee, J.; Kim, I.; Yoo, E.; Baek, S.J. Competitive inhibition by NAG-1/GDF-15 NLS peptide enhances its anti-cancer activity. Biochem. Biophys. Res. Commun. 2019, 519, 29–34. [Google Scholar] [CrossRef]
- Vinci, M.; Box, C.; Eccles, S.A. Three-Dimensional (3D) Tumor Spheroid Invasion Assay. J. Vis. Exp. 2015, e52686. [Google Scholar] [CrossRef] [Green Version]
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Lee, J.; Moon, H.; Ku, B.; Lee, K.; Hwang, C.-Y.; Baek, S.J. Anticancer Effects of Cold Atmospheric Plasma in Canine Osteosarcoma Cells. Int. J. Mol. Sci. 2020, 21, 4556. https://doi.org/10.3390/ijms21124556
Lee J, Moon H, Ku B, Lee K, Hwang C-Y, Baek SJ. Anticancer Effects of Cold Atmospheric Plasma in Canine Osteosarcoma Cells. International Journal of Molecular Sciences. 2020; 21(12):4556. https://doi.org/10.3390/ijms21124556
Chicago/Turabian StyleLee, Jaehak, Hyunjin Moon, Bonghye Ku, Keunho Lee, Cheol-Yong Hwang, and Seung Joon Baek. 2020. "Anticancer Effects of Cold Atmospheric Plasma in Canine Osteosarcoma Cells" International Journal of Molecular Sciences 21, no. 12: 4556. https://doi.org/10.3390/ijms21124556