EBNA1上调伯基特淋巴瘤细胞系p53抑制基因

IF 1.6 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Reports of Biochemistry and Molecular Biology Pub Date : 2023-01-01 DOI:10.52547/rbmb.11.4.672
Seyed Mohammad Ali Hashemi, Abdolvahab Moradi, Seyed Younes Hosseini, Hadi Razavi Nikoo, Taravat Bamdad, Mahboobeh Razmkhah, Jamal Sarvari, Alijan Tabarraei
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引用次数: 0

摘要

背景:p53的抑制是Epstein-Barr病毒相关肿瘤的重要机制,被描述为EBNA1-USP7,它是p53抑制的关键轴。因此,在本研究中,我们旨在评估EBNA1对HDAC-1、MDM2、MDM4、Sirt-3、PSMD10等p53抑制基因表达的功能,以及GNE-6776在蛋白/mRNA水平上抑制USP7对p53的影响。方法:采用电穿孔法用EBNA1转染BL28细胞系。选择EBNA1表达稳定的细胞,采用潮霉素B处理。采用实时荧光定量PCR检测PSMD10、HDAC-1、USP7、MDM2、P53、Sirt-3、MDM4等7个基因的表达。为了评估USP7的抑制效果,我们用GNE-6776处理细胞;24小时和4天后,收集细胞,再次评估感兴趣基因的表达。结果:MDM2 (P=0.028)、MDM4 (P=0.028)、USP7 (P=0.028)、HDAC1 (P=0.015)在EBNA1携带细胞中的表达均显著高于转染质粒的对照细胞,而p53 mRNA在EBNA1携带细胞中的表达仅轻微下调(P=0.685)。治疗4天后,所有研究基因都没有明显改变。治疗后24小时内p53 mRNA表达下调(P=0.685), 4 d后p53 mRNA表达上调(P=0.7),差异不显著。结论:EBNA1可能强烈上调p53抑制基因HDAC1、MDM2、MDM4和USP7。此外,USP7在蛋白/mRNA水平上对p53的抑制作用似乎与细胞性质有关;然而,还需要进一步的研究。
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EBNA1 Upregulates P53-Inhibiting Genes in Burkitt's Lymphoma Cell Line.

Background: Suppression of p53 is an important mechanism in Epstein-Barr virus associate-tumors and described as EBNA1-USP7 which is a key axis in p53 suppression. Thus, in this study, we aimed to evaluate the function of EBNA1 on the expression of p53-inhibiting genes including HDAC-1, MDM2, MDM4, Sirt-3, and PSMD10 and the influence of USP7 inhibition using GNE-6776 on p53 at protein/mRNA level.

Methods: The electroporation method was used to transfect the BL28 cell line with EBNA1. Cells with stable EBNA1 expression were selected by Hygromycin B treatment. The expression of seven genes, including PSMD10, HDAC-1, USP7, MDM2, P53, Sirt-3, and MDM4, was evaluated using a real-time PCR assay. For evaluating the effects of USP7 inhibition, the cells were treated with GNE-6776; after 24 hours and 4 days, the cells were collected and again expression of interest genes was evaluated.

Results: MDM2 (P=0.028), MDM4 (P=0.028), USP7 (P=0.028), and HDAC1 (P=0.015) all showed significantly higher expression in EBNA1-harboring cells compared to control plasmid transfected cells, while p53 mRNA expression was only marginally downregulated in EBNA1 harboring cells (P=0.685). Four-day after treatment, none of the studied genes was significantly changed. Also, in the first 24-hour after treatment, mRNA expression of p53 was downregulated (P=0.685), but after 4 days it was upregulated (P=0.7) insignificantly.

Conclusion: It seems that EBNA1 could strongly upregulate p53-inhibiting genes including HDAC1, MDM2, MDM4, and USP7. Moreover, it appears that the effects of USP7 suppression on p53 at protein/mRNA level depend on the cell nature; however, further research is needed.

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来源期刊
Reports of Biochemistry and Molecular Biology
Reports of Biochemistry and Molecular Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-
CiteScore
2.80
自引率
23.50%
发文量
60
审稿时长
10 weeks
期刊介绍: The Reports of Biochemistry & Molecular Biology (RBMB) is the official journal of the Varastegan Institute for Medical Sciences and is dedicated to furthering international exchange of medical and biomedical science experience and opinion and a platform for worldwide dissemination. The RBMB is a medical journal that gives special emphasis to biochemical research and molecular biology studies. The Journal invites original and review articles, short communications, reports on experiments and clinical cases, and case reports containing new insights into any aspect of biochemistry and molecular biology that are not published or being considered for publication elsewhere. Publications are accepted in the form of reports of original research, brief communications, case reports, structured reviews, editorials, commentaries, views and perspectives, letters to authors, book reviews, resources, news, and event agenda.
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