Seyed Mohammad Ali Hashemi, Abdolvahab Moradi, Seyed Younes Hosseini, Hadi Razavi Nikoo, Taravat Bamdad, Mahboobeh Razmkhah, Jamal Sarvari, Alijan Tabarraei
{"title":"EBNA1上调伯基特淋巴瘤细胞系p53抑制基因","authors":"Seyed Mohammad Ali Hashemi, Abdolvahab Moradi, Seyed Younes Hosseini, Hadi Razavi Nikoo, Taravat Bamdad, Mahboobeh Razmkhah, Jamal Sarvari, Alijan Tabarraei","doi":"10.52547/rbmb.11.4.672","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>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 <i>HDAC-1, MDM2, MDM4, Sirt-3</i>, and <i>PSMD10</i> and the influence of USP7 inhibition using GNE-6776 on p53 at protein/mRNA level.</p><p><strong>Methods: </strong>The electroporation method was used to transfect the BL28 cell line with <i>EBNA1</i>. Cells with stable <i>EBNA1</i> expression were selected by Hygromycin B treatment. The expression of seven genes, including <i>PSMD10, HDAC-1, USP7, MDM2, P53, Sirt-3</i>, and <i>MDM4</i>, 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.</p><p><strong>Results: </strong><i>MDM2</i> (P=0.028), <i>MDM4</i> (P=0.028), <i>USP7</i> (P=0.028), and <i>HDAC1</i> (P=0.015) all showed significantly higher expression in <i>EBNA1</i>-harboring cells compared to control plasmid transfected cells, while <i>p53</i> mRNA expression was only marginally downregulated in <i>EBNA1</i> 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.</p><p><strong>Conclusion: </strong>It seems that EBNA1 could strongly upregulate p53-inhibiting genes including <i>HDAC1, MDM2, MDM4</i>, and <i>USP7</i>. 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.</p>","PeriodicalId":45319,"journal":{"name":"Reports of Biochemistry and Molecular Biology","volume":null,"pages":null},"PeriodicalIF":1.6000,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10149133/pdf/rbmb-11-672.pdf","citationCount":"0","resultStr":"{\"title\":\"EBNA1 Upregulates P53-Inhibiting Genes in Burkitt's Lymphoma Cell Line.\",\"authors\":\"Seyed Mohammad Ali Hashemi, Abdolvahab Moradi, Seyed Younes Hosseini, Hadi Razavi Nikoo, Taravat Bamdad, Mahboobeh Razmkhah, Jamal Sarvari, Alijan Tabarraei\",\"doi\":\"10.52547/rbmb.11.4.672\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>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 <i>HDAC-1, MDM2, MDM4, Sirt-3</i>, and <i>PSMD10</i> and the influence of USP7 inhibition using GNE-6776 on p53 at protein/mRNA level.</p><p><strong>Methods: </strong>The electroporation method was used to transfect the BL28 cell line with <i>EBNA1</i>. Cells with stable <i>EBNA1</i> expression were selected by Hygromycin B treatment. The expression of seven genes, including <i>PSMD10, HDAC-1, USP7, MDM2, P53, Sirt-3</i>, and <i>MDM4</i>, 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.</p><p><strong>Results: </strong><i>MDM2</i> (P=0.028), <i>MDM4</i> (P=0.028), <i>USP7</i> (P=0.028), and <i>HDAC1</i> (P=0.015) all showed significantly higher expression in <i>EBNA1</i>-harboring cells compared to control plasmid transfected cells, while <i>p53</i> mRNA expression was only marginally downregulated in <i>EBNA1</i> 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.</p><p><strong>Conclusion: </strong>It seems that EBNA1 could strongly upregulate p53-inhibiting genes including <i>HDAC1, MDM2, MDM4</i>, and <i>USP7</i>. 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.</p>\",\"PeriodicalId\":45319,\"journal\":{\"name\":\"Reports of Biochemistry and Molecular Biology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.6000,\"publicationDate\":\"2023-01-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10149133/pdf/rbmb-11-672.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Reports of Biochemistry and Molecular Biology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.52547/rbmb.11.4.672\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Reports of Biochemistry and Molecular Biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.52547/rbmb.11.4.672","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
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.
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.