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Isolation and Quantification of Epstein-Barr Virus from the P3HR1 Cell Line
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Epstein-Barr Virus miR-BART1-3p Regulates the miR-17-92 Cluster by Targeting E2F3
Myung Chan Park1, Hyoji Kim1, Hoyun Choi1
1Department of Medical Life Sciences, Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 06591, Korea.
International Journal of Molecular Sciences
|October 23, 2021
Summary
Epstein-Barr virus (EBV) microRNA miR-BART1-3p inhibits gastric carcinoma cell growth by targeting E2F3. This suppression also affects the miR-17-92 cluster, suggesting a role in cell cycle regulation.
Area of Science:
- Oncology
- Virology
- Molecular Biology
Background:
- Epstein-Barr virus (EBV) is linked to various tumors, producing BamHI A rightward transcript (BART) microRNAs (miRNAs).
- BART miRNAs are more prevalent in EBV-associated epithelial cancers than in EBV-infected B lymphomas.
- The specific role of EBV miRNAs in cancer progression, particularly gastric carcinoma, requires further elucidation.
Purpose of the Study:
- To investigate the impact of EBV-encoded miR-BART1-3p on gastric carcinoma cell cycle and growth.
- To identify the molecular targets of miR-BART1-3p involved in cell cycle regulation.
- To explore the relationship between miR-BART1-3p, E2F3, and the miR-17-92 cluster in gastric cancer.
Main Methods:
- Transfection of miR-BART1-3p into gastric carcinoma cells.
- Microarray analysis to identify differentially expressed genes.
- Luciferase reporter assays to confirm direct targeting of E2F3 mRNA.
- Quantitative real-time PCR and Western blotting to assess gene and protein expression.
- Analysis of patient samples (EBV-associated vs. EBV-negative gastric carcinoma).
Main Results:
- miR-BART1-3p induced G0/G1 cell cycle arrest and suppressed gastric carcinoma cell growth.
- miR-BART1-3p directly targets the 3'-UTR of E2F3 mRNA, reducing its expression.
- E2F3, MIR17HG, and miR-17-92 cluster miRNAs were downregulated in EBV-associated gastric carcinoma patients.
- miR-BART1-3p and E2F3 siRNA inhibited miR-17-92 cluster expression, while miR-BART1-3p inhibition enhanced it.
Conclusions:
- miR-BART1-3p plays a significant role in inhibiting gastric carcinoma cell proliferation.
- The study identifies E2F3 as a direct target of miR-BART1-3p, mediating its effects on cell cycle progression.
- miR-BART1-3p influences the expression of the evolutionarily conserved miR-17-92 cluster via E2F3 suppression, highlighting a novel regulatory pathway in gastric cancer.
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