Cytotoxic Drugs Activate KSHV Lytic Cycle in Latently Infected PEL Cells by Inducing a Moderate ROS Increase Controlled by HSF1, NRF2 and p62/SQSTM1
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Cell Treatments
2.3. Cell Assay Viability
2.4. Antibodies
2.5. Western Blot Analysis
2.6. Indirect Immunofluorescence Assay (IFA)
2.7. Endogenous Reactive Oxygen Species (ROS) Detection
2.8. Transfection and Plasmids
2.9. HSF1, NRF2 and P62/SQSTM1 Knockdown by Small Interfering RNA (siRNA)
2.10. RNA Extraction
2.11. Reverse-Transcription Quantitative PCR (qRT-PCR) and Quantitative PCR (qPCR)
2.12. Densitometric Analysis
2.13. Statistical Analysis
3. Results
3.1. TPA/Butyrate (TB) and Bortezomib (BZ) Increase Intracellular ROS and Promote KSHV Lytic Antigen Expression, while Quercetin (Q) and Metformin (MET) Reduce ROS and Do Not Induce This Effect in PEL Cells
3.2. p62/SQSTM1 is Up-Regulated by TB and BZ and Down-Regulated By MET And Q
3.3. p62/SQSTM1 Up-Regulation is not Dependent on Autophagy Reduction at 6 and 12 h, while Autophagy Inhibition Contributes to Its Accumulation at 24 h after TB or BZ Treatments
3.4. p62/SQSTM1 Plays a Role in KSHV Lytic Antigen Expression Induced by TB or BZ
3.5. The Main Mediators of Cell Stress Responses, HSF1 and NRF2, Promote P62/SQSTM1 And KSHV Lytic Antigen Expression in TB- or BZ-Treated PEL Cells
3.6. The Inhibition of HSF1, NRF2 and p62/SQSTM1 (siRNA) Further Increases ROS and Cytotoxicity Induced by TB or BZ in PEL Cells
4. Discussion
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Granato, M.; Gilardini Montani, M.S.; Angiolillo, C.; D’Orazi, G.; Faggioni, A.; Cirone, M. Cytotoxic Drugs Activate KSHV Lytic Cycle in Latently Infected PEL Cells by Inducing a Moderate ROS Increase Controlled by HSF1, NRF2 and p62/SQSTM1. Viruses 2019, 11, 8. https://doi.org/10.3390/v11010008
Granato M, Gilardini Montani MS, Angiolillo C, D’Orazi G, Faggioni A, Cirone M. Cytotoxic Drugs Activate KSHV Lytic Cycle in Latently Infected PEL Cells by Inducing a Moderate ROS Increase Controlled by HSF1, NRF2 and p62/SQSTM1. Viruses. 2019; 11(1):8. https://doi.org/10.3390/v11010008
Chicago/Turabian StyleGranato, Marisa, Maria Saveria Gilardini Montani, Camilla Angiolillo, Gabriella D’Orazi, Alberto Faggioni, and Mara Cirone. 2019. "Cytotoxic Drugs Activate KSHV Lytic Cycle in Latently Infected PEL Cells by Inducing a Moderate ROS Increase Controlled by HSF1, NRF2 and p62/SQSTM1" Viruses 11, no. 1: 8. https://doi.org/10.3390/v11010008
APA StyleGranato, M., Gilardini Montani, M. S., Angiolillo, C., D’Orazi, G., Faggioni, A., & Cirone, M. (2019). Cytotoxic Drugs Activate KSHV Lytic Cycle in Latently Infected PEL Cells by Inducing a Moderate ROS Increase Controlled by HSF1, NRF2 and p62/SQSTM1. Viruses, 11(1), 8. https://doi.org/10.3390/v11010008