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Research
RNA helicase DHX15 decreases cell apoptosis by NF-κB signaling pathway in Burkitt lymphoma
Shanyuan Wang
Corresponding Author
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Background DHX15 is one of the RNA helicase family members involving in several biological processes, especially in anti-virus and pre-mRNA splicing. Studies have reported that overexpression of DHX15 is related to cancer progression. However, the role of DHX15 in Burkitt lymphoma (BL) and latent EB virus infection remains to be elucidated.
Methods Expression of DHX15 was measured in BL patient by immunohistochemical staining. In vitro study, a CCK-8 assay was used to analyze the proliferation of Raji cells infected with DHX15-shRNA-Lentivirus and flow cytometry was performed to assess Raji cell cycle, apoptosis and mitochondria membrane potential. Members of NF-κB signaling pathway and apoptotic-related proteins expression was measured by western-blot. EBV latent infection products and RNA polymerase Ⅲ transcripts expression were determined by quantitative real-time PCR and western-blot. In vivo study, HE, IHC, TUNEL and ISH assays were used to analyze the effect of DHX15 on subcutaneous tumor nodes formation.
Results DHX15 was overexpressed in Burkitt lymphoma patients and tends to be associated with poor progression-free survival and poor overall survival. Knockdown of DHX15 significantly inhibits BL tumor growth, reduced cell proliferation, induced G2/M-phase arrest and increased cell apoptosis. Further analysis showed that canonical NF-κB signaling and downstream targets (such as Bcl-2, Bcl-xl, survivin), mitochondria and Caspase were involved in the increased cell apoptosis after DHX15 gene knockdown. Furthermore, knockdown of DHX15 induced EBV latent infection products expression and inhibited RNA polymerase Ⅲ activity.
Conclusion DHX15 may be an oncogene in the development of BL and a potential therapeutic target for the treatment of BL and latent EBV infection.
Keywords
DHX15, Burkitt lymphoma, gene knockdown, apoptosis
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This is a preprint. It has not completed peer review.
Research
RNA helicase DHX15 decreases cell apoptosis by NF-κB signaling pathway in Burkitt lymphoma
Shanyuan Wang
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 16 Apr, 2020
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Oncology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background DHX15 is one of the RNA helicase family members involving in several biological processes, especially in anti-virus and pre-mRNA splicing. Studies have reported that overexpression of DHX15 is related to cancer progression. However, the role of DHX15 in Burkitt lymphoma (BL) and latent EB virus infection remains to be elucidated.
Methods Expression of DHX15 was measured in BL patient by immunohistochemical staining. In vitro study, a CCK-8 assay was used to analyze the proliferation of Raji cells infected with DHX15-shRNA-Lentivirus and flow cytometry was performed to assess Raji cell cycle, apoptosis and mitochondria membrane potential. Members of NF-κB signaling pathway and apoptotic-related proteins expression was measured by western-blot. EBV latent infection products and RNA polymerase Ⅲ transcripts expression were determined by quantitative real-time PCR and western-blot. In vivo study, HE, IHC, TUNEL and ISH assays were used to analyze the effect of DHX15 on subcutaneous tumor nodes formation.
Results DHX15 was overexpressed in Burkitt lymphoma patients and tends to be associated with poor progression-free survival and poor overall survival. Knockdown of DHX15 significantly inhibits BL tumor growth, reduced cell proliferation, induced G2/M-phase arrest and increased cell apoptosis. Further analysis showed that canonical NF-κB signaling and downstream targets (such as Bcl-2, Bcl-xl, survivin), mitochondria and Caspase were involved in the increased cell apoptosis after DHX15 gene knockdown. Furthermore, knockdown of DHX15 induced EBV latent infection products expression and inhibited RNA polymerase Ⅲ activity.
Conclusion DHX15 may be an oncogene in the development of BL and a potential therapeutic target for the treatment of BL and latent EBV infection.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6