São Paulo State University (UNESP), Department of Pathology, Botucatu Medical School, Av. Prof. Dr. Mário Rubens Guimarães Montenegro S/n, CEP 18618-687, Botucatu, São Paulo, Brazil; São Paulo State University (UNESP), Institute of Biotechnology (IBTEC), Alameda Das Tecomarias S/n, CEP 18607-440, Botucatu, São Paulo, Brazil.
São Paulo State University (UNESP), Institute of Biotechnology (IBTEC), Alameda Das Tecomarias S/n, CEP 18607-440, Botucatu, São Paulo, Brazil; São Paulo State University (UNESP), Institute of Biosciences (IBB). R. Prof. Dr. Antônio Celso Wagner Zanin, 250, CEP 18618-689, Botucatu, São Paulo, Brazil.
Tumour Virus Res. 2024 Jun;17:200276. doi: 10.1016/j.tvr.2023.200276. Epub 2023 Dec 29.
The Epstein-Barr Virus (EBV) encodes viral microRNAs (miRs) that have been implicated in the pathogenesis of nasopharyngeal and gastric carcinomas, yet their potential roles in lymphomas remain to be fully elucidated. This study evaluated the impact of CRISPR/Cas9-mediated knockdown of EBV miRs BART-7 and BART-9 in EBV-positive Burkitt lymphoma cells Akata. As anticipated, the Akata cells subjected to CRISPR/Cas9-mediated knockdown of either EBV BART-7 or BART-9 exhibited a significant reduction in the expression of these viral miRs compared to cells with wild-type (wt) EBV genomes. This outcome effectively validates the experimental model employed in this study. Knocking down either BART-7 or BART-9 resulted in a notable reduction in cell viability and proliferation rates, alongside an elevation in the expression of EBV lytic genes. Global proteomic analysis revealed that the knockdown of EBV BART-7 significantly decreased the expression of ubiquitin/proteasome proteins while concurrently increasing RNA binding proteins (RBPs). Conversely, BART-9 knockdown reduced proteins associated with oxidoreductase activity, particularly those involved in fatty acid metabolism. Our findings unveil previously undiscovered EBV miRs BARTs 7 and 9 roles in cellular pathways relevant to both viral biology and lymphomagenesis.
EBV 编码的病毒 microRNAs(miRs)已被认为与鼻咽癌和胃癌的发病机制有关,但它们在淋巴瘤中的潜在作用仍有待充分阐明。本研究评估了 CRISPR/Cas9 介导的 EBV 微小 RNA BART-7 和 BART-9 敲低对 EBV 阳性 Burkitt 淋巴瘤细胞 Akata 的影响。正如预期的那样,与具有野生型(wt) EBV 基因组的细胞相比,经 CRISPR/Cas9 介导的 EBV BART-7 或 BART-9 敲低的 Akata 细胞中这些病毒微小 RNA 的表达显著降低。这一结果有效地验证了本研究中使用的实验模型。敲低 BART-7 或 BART-9 均可显著降低细胞活力和增殖率,并上调 EBV 裂解基因的表达。全蛋白质组分析显示,EBV BART-7 的敲低显著降低了泛素/蛋白酶体蛋白的表达,同时增加了 RNA 结合蛋白(RBPs)的表达。相反,BART-9 的敲低降低了与氧化还原酶活性相关的蛋白,特别是与脂肪酸代谢相关的蛋白。我们的发现揭示了以前未发现的 EBV 微小 RNA BARTs7 和 9 在与病毒生物学和淋巴瘤发生相关的细胞途径中的作用。