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Volume 11, Issue 4 (Vol.11 No.4 Jan 2023)
rbmb.net 2023, 11(4): 672-683 |
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Hashemi S M A, Moradi A, Hosseini S Y, Razavi Nikoo H, Bamdad T, Razmkhah M, et al . EBNA1 Upregulates P53-Inhibiting Genes in Burkitt's Lymphoma Cell Line. rbmb.net 2023; 11 (4) :672-683
URL: http://rbmb.net/article-1-1012-en.html
URL: http://rbmb.net/article-1-1012-en.html
Seyed Mohammad Ali Hashemi 
, Abdolvahab Moradi , Seyed Younes Hosseini , Hadi Razavi Nikoo , Taravat Bamdad , Mahboobeh Razmkhah , Jamal Sarvari * , Alijan Tabarraei*
, Abdolvahab Moradi , Seyed Younes Hosseini , Hadi Razavi Nikoo , Taravat Bamdad , Mahboobeh Razmkhah , Jamal Sarvari * , Alijan Tabarraei*
Department of Bacteriology and Virology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran & Gastroenterohepatology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Abstract: (1978 Views)
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.
Conclusions: 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.
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.
Conclusions: 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.
Type of Article: Original Article |
Subject:
Microbiology
Received: 2022/07/29 | Accepted: 2022/08/14 | Published: 2023/04/3
Received: 2022/07/29 | Accepted: 2022/08/14 | Published: 2023/04/3
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