Intracellular Iron Chelation by a Novel Compound, C7, Reactivates Epstein–Barr Virus (EBV) Lytic Cycle via the ERK-Autophagy Axis in EBV-Positive Epithelial Cancers
Abstract
:1. Introduction
2. Results
2.1. Effects of C7 and Its Analog Compounds on EBV Lytic Cycle Reactivation
2.2. Binding of C7 and Its Lytic-Inducing Analogs to Iron
2.3. EBV Lytic Reactivation by Other Clinically Available Iron Chelators
2.4. Iron Chelation by C7 and Other Iron Chelators Is Required for EBV Lytic Reactivation
2.5. Enrichment of the Hallmark Gene Set in Hypoxia Signaling Pathway by C7
2.6. EBV Lytic Reactivation by C7 Requires the Activation of ERK in Addition to Hypoxia Signaling Pathway
2.7. Intracellular Iron Chelation Leads to the Induction of the ERK-Autophagy Axis and Reactivation of EBV Lytic Cycle
2.8. C7 Sensitizes EBV-Positive Gastric Carcinoma Cells to Killing by Ganciclovir
3. Discussion
4. Materials and Methods
4.1. Cell Lines and Culturing Conditions
4.2. Drug Treatment
4.3. Calcein-AM Measurement
4.4. Western Blot Analysis
4.5. RNA-Sequencing Analysis
4.6. Bioinformatics Analysis
4.7. Metal Binding Titration, Measurement, and Analysis
4.8. Immunofluorescence Staining
4.9. MTT Assay
4.10. CRISPR Gene Knockout
4.11. Transient Gene Knockdown
4.12. Quantitative PCR Assay
4.13. Luciferase Reporter Assay
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Yiu, S.P.T.; Hui, K.F.; Choi, C.K.; Kao, R.Y.T.; Ma, C.W.; Yang, D.; Chiang, A.K.S. Intracellular Iron Chelation by a Novel Compound, C7, Reactivates Epstein–Barr Virus (EBV) Lytic Cycle via the ERK-Autophagy Axis in EBV-Positive Epithelial Cancers. Cancers 2018, 10, 505. https://doi.org/10.3390/cancers10120505
Yiu SPT, Hui KF, Choi CK, Kao RYT, Ma CW, Yang D, Chiang AKS. Intracellular Iron Chelation by a Novel Compound, C7, Reactivates Epstein–Barr Virus (EBV) Lytic Cycle via the ERK-Autophagy Axis in EBV-Positive Epithelial Cancers. Cancers. 2018; 10(12):505. https://doi.org/10.3390/cancers10120505
Chicago/Turabian StyleYiu, Stephanie Pei Tung, Kwai Fung Hui, Chung King Choi, Richard Yi Tsun Kao, Chi Wang Ma, Dan Yang, and Alan Kwok Shing Chiang. 2018. "Intracellular Iron Chelation by a Novel Compound, C7, Reactivates Epstein–Barr Virus (EBV) Lytic Cycle via the ERK-Autophagy Axis in EBV-Positive Epithelial Cancers" Cancers 10, no. 12: 505. https://doi.org/10.3390/cancers10120505