Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cells
Abstract Background Epstein-Barr Virus (EBV) is associated with hematopoietic malignancies, such as Burkitt’s lymphoma, post-transplantation lymphoproliferative disorder, and diffuse large B-cell lymphoma. The current approach for EBV-associated lymphoma involves chemotherapy to eradicate cancer cel...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-06-01
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| Series: | Virology Journal |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12985-017-0784-7 |
| _version_ | 1818153575984922624 |
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| author | Qinyan Yin Mark Sides Christopher H. Parsons Erik K. Flemington Joseph A. Lasky |
| author_facet | Qinyan Yin Mark Sides Christopher H. Parsons Erik K. Flemington Joseph A. Lasky |
| author_sort | Qinyan Yin |
| collection | DOAJ |
| description | Abstract Background Epstein-Barr Virus (EBV) is associated with hematopoietic malignancies, such as Burkitt’s lymphoma, post-transplantation lymphoproliferative disorder, and diffuse large B-cell lymphoma. The current approach for EBV-associated lymphoma involves chemotherapy to eradicate cancer cells, however, normal cells may be injured and organ dysfunction may occur with currently employed regimens. This research is focused on employing arsenic trioxide (ATO) as EBV-specific cancer therapy takes advantage of the fact the EBV resides within the malignant cells. Methods and results Our research reveals that low ATO inhibits EBV gene expression and genome replication. EBV spontaneous reactivation starts as early as 6 h after re-suspending EBV-positive Mutu cells in RPMI media in the absence of ATO, however this does not occur in Mutu cells cultured with ATO. ATO’s inhibition of EBV spontaneous reactivation is dose dependent. The expression of the EBV immediate early gene Zta and early gene BMRF1 is blocked with low concentrations of ATO (0.5 nM – 2 nM) in EBV latency type I cells and EBV-infected PBMC cells. The combination of ATO and ganciclovir further diminishes EBV gene expression. ATO-mediated reduction of EBV gene expression can be rescued by co-treatment with the proteasome inhibitor MG132, indicating that ATO promotes ubiquitin conjugation and proteasomal degradation of EBV genes. Co-immunoprecipitation assays with antibodies against Zta pulls down more ubiquitin in ATO treated cell lysates. Furthermore, MG132 reverses the inhibitory effect of ATO on anti-IgM-, PMA- and TGF-β-mediated EBV reactivation. Thus, mechanistically ATO’s inhibition of EBV gene expression occurs via the ubiquitin pathway. Moreover, ATO treatment results in increased cell death in EBV-positive cells compared to EBV-negative cells, as demonstrated by both MTT and trypan blue assays. ATO-induced cell death in EBV-positive cells is dose dependent. ATO and ganciclovir in combination further enhances cell death specifically in EBV-positive cells. Conclusion ATO-mediated inhibition of EBV lytic gene expression results in cell death selectively in EBV-positive lymphocytes, suggesting that ATO may potentially serve as a drug to treat EBV-related lymphomas in the clinical setting. |
| first_indexed | 2024-12-11T14:12:48Z |
| format | Article |
| id | doaj.art-6a07a2f0d1cd40fdb8d48a5fb3ce3dbe |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-11T14:12:48Z |
| publishDate | 2017-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-6a07a2f0d1cd40fdb8d48a5fb3ce3dbe2022-12-22T01:03:20ZengBMCVirology Journal1743-422X2017-06-0114111210.1186/s12985-017-0784-7Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cellsQinyan Yin0Mark Sides1Christopher H. Parsons2Erik K. Flemington3Joseph A. Lasky4Department of Medicine, Section of Pulmonary Disease, Tulane University School of MedicineDepartment of Medicine, Section of Pulmonary Disease, Tulane University School of MedicineDepartment of Internal Medicine, Louisiana University School of MedicineDepartment of Pathology and Laboratory, Tulane University School of MedicineDepartment of Medicine, Section of Pulmonary Disease, Tulane University School of MedicineAbstract Background Epstein-Barr Virus (EBV) is associated with hematopoietic malignancies, such as Burkitt’s lymphoma, post-transplantation lymphoproliferative disorder, and diffuse large B-cell lymphoma. The current approach for EBV-associated lymphoma involves chemotherapy to eradicate cancer cells, however, normal cells may be injured and organ dysfunction may occur with currently employed regimens. This research is focused on employing arsenic trioxide (ATO) as EBV-specific cancer therapy takes advantage of the fact the EBV resides within the malignant cells. Methods and results Our research reveals that low ATO inhibits EBV gene expression and genome replication. EBV spontaneous reactivation starts as early as 6 h after re-suspending EBV-positive Mutu cells in RPMI media in the absence of ATO, however this does not occur in Mutu cells cultured with ATO. ATO’s inhibition of EBV spontaneous reactivation is dose dependent. The expression of the EBV immediate early gene Zta and early gene BMRF1 is blocked with low concentrations of ATO (0.5 nM – 2 nM) in EBV latency type I cells and EBV-infected PBMC cells. The combination of ATO and ganciclovir further diminishes EBV gene expression. ATO-mediated reduction of EBV gene expression can be rescued by co-treatment with the proteasome inhibitor MG132, indicating that ATO promotes ubiquitin conjugation and proteasomal degradation of EBV genes. Co-immunoprecipitation assays with antibodies against Zta pulls down more ubiquitin in ATO treated cell lysates. Furthermore, MG132 reverses the inhibitory effect of ATO on anti-IgM-, PMA- and TGF-β-mediated EBV reactivation. Thus, mechanistically ATO’s inhibition of EBV gene expression occurs via the ubiquitin pathway. Moreover, ATO treatment results in increased cell death in EBV-positive cells compared to EBV-negative cells, as demonstrated by both MTT and trypan blue assays. ATO-induced cell death in EBV-positive cells is dose dependent. ATO and ganciclovir in combination further enhances cell death specifically in EBV-positive cells. Conclusion ATO-mediated inhibition of EBV lytic gene expression results in cell death selectively in EBV-positive lymphocytes, suggesting that ATO may potentially serve as a drug to treat EBV-related lymphomas in the clinical setting.http://link.springer.com/article/10.1186/s12985-017-0784-7Epstein-Barr virusEBVArsenic trioxideATOLymphomaCancer |
| spellingShingle | Qinyan Yin Mark Sides Christopher H. Parsons Erik K. Flemington Joseph A. Lasky Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cells Virology Journal Epstein-Barr virus EBV Arsenic trioxide ATO Lymphoma Cancer |
| title | Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cells |
| title_full | Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cells |
| title_fullStr | Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cells |
| title_full_unstemmed | Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cells |
| title_short | Arsenic trioxide inhibits EBV reactivation and promotes cell death in EBV-positive lymphoma cells |
| title_sort | arsenic trioxide inhibits ebv reactivation and promotes cell death in ebv positive lymphoma cells |
| topic | Epstein-Barr virus EBV Arsenic trioxide ATO Lymphoma Cancer |
| url | http://link.springer.com/article/10.1186/s12985-017-0784-7 |
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