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Updated: Aug 18, 2026
Isolation and Quantification of Epstein-Barr Virus from the P3HR1 Cell Line
Published on: September 28, 2022
Effect of EBV LMP1 targeted DNAzymes on cell proliferation and apoptosis
Zhong-Xin Lu1, Mao Ye, Guang-Rong Yan
1Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, China.
Abstract:
The latent membrane protein (LMP1) encoded by Epstein-Barr virus (EBV) has been suggested to be one of the major oncogenic factors in EBV-mediated carcinogenesis. RNA-cleaving DNA enzymes are catalytic nucleic acids that bind and cleave a target RNA in a highly sequence-specific manner. In this study, we explore the potential of using DNAzymes as a therapeutic approach to EBV-associated carcinomas by targeting the LMP1 gene. In all, 13 different phosphorothioate-modified "10-23" deoxyribozymes (DNAzymes) were designed and synthesized against the LMP1 mRNA and transfected into B95-8 cells, which constitutively express the LMP1. Fluorescence microscopy was used to examine the cellular uptake and distribution in B95-8 cells. As demonstrated in Western blots, three out of 13 deoxyribozymes significantly downregulated the expression of LMP1 in B95-8 cells. These DNAzymes were shown to markedly inhibit B95-8 cell growth compared with a disabled DNAzyme and untreated controls, as determined by an alamarBlue Assay. It was further demonstrated that these DNAzymes arrested the B95-8 cells in G0/G1 using flow cytometry. Interestingly, the active DNAzymes could also downregulate the expression of Bcl-2 gene in treated cells, suggesting a close association between the LMP1 and Bcl-2 genes and their involvement in apoptosis. This was further confirmed with the result that the DNAzymes could induce the release of cytochrome c from mitochondria, which is the hallmark of the apoptosis. The present results suggest that the LMP1 may present a potential target for DNAzymes towards the EBV-associated carcinoma through cell proliferation and apoptosis pathways.
Insights
DNAzymes targeting Epstein-Barr virus latent membrane protein 1 (LMP1) significantly inhibited cancer cell growth and induced apoptosis. These findings highlight DNAzymes as a promising therapeutic strategy for EBV-associated carcinomas.
Area of Science:
- Molecular Biology
- Cancer Research
- Biochemistry
Background:
- Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) is a key oncogenic factor in EBV-mediated carcinogenesis.
- RNA-cleaving DNAzymes offer sequence-specific targeting of RNA molecules.
- Targeting LMP1 presents a potential therapeutic strategy for EBV-associated cancers.
Purpose of the Study:
- To investigate the therapeutic potential of DNAzymes against EBV LMP1 mRNA in EBV-associated carcinomas.
- To evaluate the efficacy of DNAzymes in downregulating LMP1 expression and inhibiting cancer cell proliferation.
- To explore the impact of DNAzymes on apoptosis pathways in EBV-infected cells.
Main Methods:
- Design and synthesis of 13 phosphorothioate-modified "10-23" DNAzymes targeting LMP1 mRNA.
- Transfection of DNAzymes into EBV-producing B95-8 cells.
- Assessment of cellular uptake, LMP1 and Bcl-2 expression (Western blot), cell viability (alamarBlue Assay), cell cycle arrest (flow cytometry), and cytochrome c release.
Main Results:
- Three out of 13 DNAzymes significantly downregulated LMP1 expression in B95-8 cells.
- Active DNAzymes markedly inhibited B95-8 cell growth and induced G0/G1 cell cycle arrest.
- DNAzymes downregulated Bcl-2 expression and induced cytochrome c release, indicating apoptosis induction.
Conclusions:
- DNAzymes targeting LMP1 are effective in reducing EBV-infected cell proliferation and inducing apoptosis.
- LMP1 and Bcl-2 are closely associated in EBV-mediated apoptosis pathways.
- DNAzymes targeting LMP1 represent a promising therapeutic approach for EBV-associated carcinomas.
