Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethics Statement
2.2. Mammalian Cell Culture
2.3. Viruses and Virus Production
2.4. Detection of Infectious H-1PV Particles
2.5. Western Blot Analysis
2.6. Microscopy
2.7. Viral DNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (qPCR)
2.8. Cell Viability and Cell Death Assessment
2.9. Flow Cytometric Characterization of the Cell-Cycle Distribution of Cells and of the Sub-G1 Apoptotic Cell Population
3. Results
3.1. Protoparvovirus H-1PV Enters and Transduces Non-Transformed and Transformed Mesenchymal Cells
3.2. H-1PV Infection of Non-Transformed Mesenchymal Cells Induces Antiproliferative Effects and Toxicity Only at High Virus Doses
3.3. H-1PV Viral Protein Expression Induces Cell Cycle Arrest in G2/M in Osteosarcoma Cells
3.4. Osteosarcoma Cells Undergo Lytic H-1PV Infection
3.5. In Osteosarcoma Cells, H-1PV Infection Does Not Lead to Efficient Production of Infectious Viral Progeny
3.6. Wild-Type H-1PV Is More Cytotoxic Towards Osteosarcoma Cells Than H-1PV-Derived Fitness Mutants
4. Discussion
4.1. Safety Profile of Wild-Type H-1PV in Human Mesenchymal Cells
4.2. Osteosarcoma Cells Are Semi-Permissive to Wild Type H-1PV Infection
4.3. Fitness Mutants Do Not Show Increased Cytotoxicity Towards Osteosarcoma Cells In Vitro
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Cell Culture | Del H-1PV | DM H-1PV | wt H-1PV |
---|---|---|---|
CAL-72 | 10 | 1 | 0.5 |
H-OS | 5 | 5 | 1 |
MG-63 | n. r. | 25 | 10 |
SaOS-2 | >10 | 10 | 5 |
U2-OS | n. r. | 5 | 5 |
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Geiss, C.; Kis, Z.; Leuchs, B.; Frank-Stöhr, M.; Schlehofer, J.R.; Rommelaere, J.; Dinsart, C.; Lacroix, J. Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro. Viruses 2017, 9, 301. https://doi.org/10.3390/v9100301
Geiss C, Kis Z, Leuchs B, Frank-Stöhr M, Schlehofer JR, Rommelaere J, Dinsart C, Lacroix J. Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro. Viruses. 2017; 9(10):301. https://doi.org/10.3390/v9100301
Chicago/Turabian StyleGeiss, Carsten, Zoltán Kis, Barbara Leuchs, Monika Frank-Stöhr, Jörg R. Schlehofer, Jean Rommelaere, Christiane Dinsart, and Jeannine Lacroix. 2017. "Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro" Viruses 9, no. 10: 301. https://doi.org/10.3390/v9100301
APA StyleGeiss, C., Kis, Z., Leuchs, B., Frank-Stöhr, M., Schlehofer, J. R., Rommelaere, J., Dinsart, C., & Lacroix, J. (2017). Preclinical Testing of an Oncolytic Parvovirus: Standard Protoparvovirus H-1PV Efficiently Induces Osteosarcoma Cell Lysis In Vitro. Viruses, 9(10), 301. https://doi.org/10.3390/v9100301