{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T22:24:32Z","timestamp":1790720672242,"version":"4.1.0"},"reference-count":56,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,10,1]],"date-time":"2024-10-01T00:00:00Z","timestamp":1727740800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>High-risk human papillomavirus (HPV) is a carcinogenic virus associated with nearly all cases of cervical cancer, as well as an increasing number of anal and oral cancers. The two carcinogenic proteins of HPV, E6 and E7, can immortalize keratinocytes and are essential for HPV-related cellular transformation. Currently, the global regulatory effects of these oncogenic proteins on the host proteome are not fully understood, and further exploration of the functions and carcinogenic mechanisms of E6 and E7 proteins is needed.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We used a previously established platform in our laboratory for constructing recombinant adenoviral plasmids expressing the HPV16 E7 gene to further construct recombinant virus particles expressing HPV16\/18 E6, E7, and both E6 and E7 genes. These recombinant viruses were used to infect C33A cells to achieve sustained expression of the HPV16\/18 E6\/E7 genes. Subsequently, total RNA was extracted and RNA-Seq technology was employed for transcriptome sequencing to identify differentially expressed genes associated with HPV infection in cervical cancer.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>RNA-Seq analysis revealed that overexpression of the HPV16\/18 E6\/E7 genes upregulated GP6, CD36, HDAC6, ESPL1, and DNMT3B among the differentially expressed genes (DEGs) associated with cervical cancer. Spearman correlation analysis revealed a statistically significant correlation between the HDAC6 and DNMT3B genes and key pathways, including DNA replication, tumor proliferation signature, G2M checkpoint, p53 pathways, and PI3K\/AKT\/mTOR signaling pathways. Further, qRT-PCR and Western blot analyses indicated that both HPV16\/18 E7 can upregulate the expression of HDAC6 and DNMT3B, genes associated with HPV infection-related cervical cancer.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The successful expression of HPV16\/18 E6\/E7 in cells indicates that the recombinant viruses retain the replication and infection capabilities of Ad4. Furthermore, the recombinant viruses expressing HPV16\/18 E7 can upregulate the HDAC6 and DNMT3B genes involved in cervical cancer pathways, thereby influencing the cell cycle. Additionally, HDAC6 and DNMT3B are emerging as important therapeutic targets for cancer. This study lays the foundation for further exploration of the oncogenic mechanisms of HPV E6\/E7 and may provide new directions for the treatment of HPV-related cancers.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2024.1459572","type":"journal-article","created":{"date-parts":[[2024,10,1]],"date-time":"2024-10-01T05:10:35Z","timestamp":1727759435000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Recombinant adenoviruses expressing HPV16\/18 E7 upregulate the HDAC6 and DNMT3B genes in C33A cells"],"prefix":"10.3389","volume":"14","author":[{"given":"Yunting","family":"Shao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pir Tariq","family":"Shah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qisheng","family":"Su","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shanhu","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fang","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chengjun","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,10,1]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1016\/j.ccr.2008.10.007","article-title":"Inactivation of the CYLD deubiquitinase by HPV E6 mediates hypoxia-induced NF-kappaB activation","volume":"14","author":"An","year":"2008","journal-title":"Cancer Cell"},{"key":"B2","doi-asserted-by":"publisher","first-page":"E11138","DOI":"10.1073\/pnas.1801156115","article-title":"Vorinostat, a pan-HDAC inhibitor, abrogates productive HPV-18 DNA amplification","volume":"115","author":"Banerjee","year":"2018","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B3","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1186\/s12885-022-09262-4","article-title":"Transcriptome analysis of cervical cancer exosomes and detection of HPVE6*I transcripts in exosomal RNA","volume":"22","author":"Bhat","year":"2022","journal-title":"BMC Cancer"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006846","article-title":"A new cell culture model to genetically dissect the complete human papillomavirus life cycle","volume":"14","author":"Bienkowska-Haba","year":"2018","journal-title":"PloS Pathog."},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.3390\/ijms20092188","article-title":"PI3K\/AKT\/mTOR signaling regulates the virus\/host cell crosstalk in HPV-positive cervical cancer cells","volume":"20","author":"Bossler","year":"2019","journal-title":"Int. J. Mol. Sci."},{"key":"B6","doi-asserted-by":"publisher","first-page":"2259","DOI":"10.3892\/ol.2017.7564","article-title":"PD-L1 expression is associated with p16(INK4A) expression in non-oropharyngeal head and neck squamous cell carcinoma","volume":"15","author":"Chen","year":"2018","journal-title":"Oncol. Lett."},{"key":"B7","doi-asserted-by":"publisher","first-page":"2438","DOI":"10.1101\/gad.10.19.2438","article-title":"p53 levels, functional domains, and DNA damage determine the extent of the apoptotic response of tumor cells","volume":"10","author":"Chen","year":"1996","journal-title":"Genes Dev."},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21207564","article-title":"Using a human papillomavirus model to study DNA replication and repair of wild type and damaged DNA templates in mammalian cells","volume":"21","author":"Das","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.3390\/v14081764","article-title":"Human papillomavirus 16 E6 and E7 oncoproteins alter the abundance of proteins associated with DNA damage response, immune signaling and epidermal differentiation","volume":"14","author":"Dust","year":"2022","journal-title":"Viruses"},{"key":"B10","doi-asserted-by":"publisher","first-page":"123","DOI":"10.3892\/ol.2010.205","article-title":"Nordihydroguaiaretic acid inhibits growth of cervical cancer SiHa cells by up-regulating p21","volume":"2","author":"Gao","year":"2011","journal-title":"Oncol. Lett."},{"key":"B11","doi-asserted-by":"publisher","first-page":"4632","DOI":"10.3390\/cimb45060294","article-title":"Gene expression profiles of methyltransferases and demethylases associated with metastasis, tumor invasion, cpG73 methylation, and HPV status in head and neck squamous cell carcinoma","volume":"45","author":"Gori\u010dan","year":"2023","journal-title":"Curr. Issues Mol. Biol."},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0017734","article-title":"Potent anti-tumor effect generated by a novel human papillomavirus (HPV) antagonist peptide reactivating the pRb\/E2F pathway","volume":"6","author":"Guo","year":"2011","journal-title":"PloS One"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.3390\/ijms24097729","article-title":"Effect of ethanol-induced methyl donors consumption on the state of hypomethylation in cervical cancer","volume":"24","author":"Han","year":"2023","journal-title":"Int. J. Mol. Sci."},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.3390\/pharmacy5030045","article-title":"Pharmacists\u2019 Attitudes and perceived barriers to human papillomavirus (HPV) vaccination services","volume":"5","author":"Hastings","year":"2017","journal-title":"Pharm. (Basel Switzerland)"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms9122496","article-title":"Epstein-barr virus infection alone or jointly with human papillomavirus associates with down-regulation of miR-145 in oral squamous-cell carcinoma","volume":"9","author":"Heawchaiyaphum","year":"2021","journal-title":"Microorganisms"},{"key":"B16","doi-asserted-by":"publisher","first-page":"158","DOI":"10.1016\/j.tim.2017.07.007","article-title":"The HPV E6\/E7 oncogenes: key factors for viral carcinogenesis and therapeutic targets","volume":"26","author":"Hoppe-Seyler","year":"2018","journal-title":"Trends Microbiol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1186\/s12929-014-0111-1","article-title":"Combination of proteasome and HDAC inhibitor enhances HPV16 E7-specific CD8+ T cell immune response and antitumor effects in a preclinical cervical cancer model","volume":"22","author":"Huang","year":"2015","journal-title":"J. Biomed. Sci."},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.3390\/v13101998","article-title":"Genomic signatures in HPV-associated tumors","volume":"13","author":"Hussain","year":"2021","journal-title":"Viruses"},{"key":"B19","doi-asserted-by":"publisher","first-page":"1293","DOI":"10.1080\/22221751.2022.2065932","article-title":"N-chlorotaurine is highly active against respiratory viruses including SARS-CoV-2 (COVID-19) in vitro","volume":"11","author":"Lackner","year":"2022","journal-title":"Emerg Microbes Infect."},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21124505","article-title":"Hub proteins involved in RAW 264.7 macrophages exposed to direct current electric field","volume":"21","author":"Li","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B21","doi-asserted-by":"publisher","first-page":"570","DOI":"10.1158\/1078-0432.ccr-08-1813","article-title":"Combination of proteasome and HDAC inhibitors for uterine cervical cancer treatment","volume":"15","author":"Lin","year":"2009","journal-title":"Clin. Cancer. Res."},{"key":"B22","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1111\/jcmm.14774","article-title":"Exosomes from human urine-derived stem cells enhanced neurogenesis via miR-26a\/HDAC6 axis after ischaemic stroke","volume":"24","author":"Ling","year":"2020","journal-title":"J. Cell. Mol. Med."},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3390\/cancers14194895","article-title":"HPV16 E7 nucleotide variants found in cancer-free subjects affect E7 protein expression and transformation","volume":"14","author":"Lou","year":"2022","journal-title":"Cancers (Basel)"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2020.592868","article-title":"Histone deacetylase inhibitors as therapeutic interventions on cervical cancer induced by human papillomavirus","volume":"8","author":"Louren\u00e7o de Freitas","year":"2020","journal-title":"Front. Cell Dev. Biol."},{"key":"B25","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1016\/j.canlet.2008.06.030","article-title":"Apicidin down-regulates human papillomavirus type 16 E6 and E7 transcripts and proteins in SiHa cervical cancer cells","volume":"272","author":"Luczak","year":"2008","journal-title":"Cancer Lett."},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0223658","article-title":"The cost-effectiveness of controlling cervical cancer using a new 9-valent human papillomavirus vaccine among school-aged girls in Australia","volume":"14","author":"Mahumud","year":"2019","journal-title":"PloS One"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines10040562","article-title":"A video-based behavioral intervention associated with improved HPV knowledge and intention to vaccinate","volume":"10","author":"Marshall","year":"2022","journal-title":"Vaccines"},{"key":"B28","doi-asserted-by":"publisher","first-page":"62","DOI":"10.1016\/j.cellimm.2011.04.005","article-title":"Immunization with a poly (lactide co-glycolide) encapsulated plasmid DNA expressing antigenic regions of HPV 16 and 18 results in an increase in the precursor frequency of T cells that respond to epitopes from HPV 16, 18, 6 and 11","volume":"270","author":"Matijevic","year":"2011","journal-title":"Cell Immunol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1007\/978-3-319-24738-0_4","article-title":"Epigenetic dysregulation in virus-associated neoplasms","volume":"879","author":"Minarovits","year":"2016","journal-title":"Adv. Exp. Med. Biol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"235","DOI":"10.34172\/bi.2020.30","article-title":"Classification and identification of human papillomavirus based on its prevalence and development of cervical lesion among Iranian women","volume":"10","author":"Moeinzadeh","year":"2020","journal-title":"BioImpacts: BI."},{"key":"B31","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1038\/nrc2886","article-title":"Human papillomavirus oncoproteins: pathways to transformation","volume":"10","author":"Moody","year":"2010","journal-title":"Nat. Rev. Cancer"},{"key":"B32","doi-asserted-by":"publisher","first-page":"93","DOI":"10.4103\/sajc.sajc_96_17","article-title":"Human papillomavirus in head and neck cancer in India: Current status and consensus recommendations","volume":"6","author":"Murthy","year":"2017","journal-title":"South Asian J. Cancer"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.3390\/ijms19061735","article-title":"Role of the DNA damage response in human papillomavirus RNA splicing and polyadenylation","volume":"19","author":"Nilsson","year":"2018","journal-title":"Int. J. Mol. Sci."},{"key":"B34","doi-asserted-by":"publisher","first-page":"1111","DOI":"10.1038\/s41598-020-80193-5","article-title":"Inhibition of kinase IKK\u03b2 suppresses cellular abnormalities induced by the human papillomavirus oncoprotein HPV 18E6","volume":"11","author":"Padash Barmchi","year":"2021","journal-title":"Sci. Rep."},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1042\/bsr20210670","article-title":"Elucidation of rutin\u2019s role in inducing caspase-dependent apoptosis via HPV-E6 and E7 down-regulation in cervical cancer HeLa cells","volume":"41","author":"Pandey","year":"2021","journal-title":"Biosci. Rep."},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1172\/jci.insight.141912","article-title":"Characterization of recombinant gorilla adenovirus HPV therapeutic vaccine PRGN-2009","volume":"6","author":"Pellom","year":"2021","journal-title":"JCI Insight"},{"key":"B37","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1186\/s13027-020-00312-9","article-title":"Overexpression of programmed cell death ligand 1 in patients with CIN and its correlation with human papillomavirus infection and CIN persistence","volume":"15","author":"Sancakli Usta","year":"2020","journal-title":"Infect. Agent. Cancer"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1177\/1010428317699799","article-title":"Impact of viral and host DNA methylations on HPV16-related cervical cancer pathogenesis","volume":"39","author":"Sen","year":"2017","journal-title":"Tumour Biol."},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3390\/cancers13071644","article-title":"HDAC6 signaling at primary cilia promotes proliferation and restricts differentiation of glioma cells","volume":"13","author":"Shi","year":"2021","journal-title":"Cancers (Basel)"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1006661","article-title":"KDM6A addiction of cervical carcinoma cell lines is triggered by E7 and mediated by p21CIP1 suppression of replication stress","volume":"13","author":"Soto","year":"2017","journal-title":"PloS Pathog."},{"key":"B41","doi-asserted-by":"publisher","first-page":"13404","DOI":"10.1038\/s41598-021-92875-9","article-title":"Combination immunotherapy with two attenuated Listeria strains carrying shuffled HPV-16 E6E7 protein causes tumor regression in a mouse tumor model","volume":"11","author":"Su","year":"2021","journal-title":"Sci. Rep."},{"key":"B42","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1038\/onc.2012.29","article-title":"Epigenetic silencing of PTPRR activates MAPK signaling, promotes metastasis and serves as a biomarker of invasive cervical cancer","volume":"32","author":"Su","year":"2013","journal-title":"Oncogene"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.3390\/ijerph19095191","article-title":"Effect of different educational interventions on knowledge of HPV vaccination and cervical cancer among young women: A preliminary report","volume":"19","author":"Takahashi","year":"2022","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1016\/j.tvr.2021.200213","article-title":"Human papillomavirus E6 and E7: What remains","volume":"11","author":"Vats","year":"2021","journal-title":"Tumour Virus Res."},{"key":"B45","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1186\/s13027-022-00417-3","article-title":"Construction and immune effect of an HPV16\/18\/58 trivalent therapeutic adenovirus vector vaccine","volume":"17","author":"Wan","year":"2022","journal-title":"Infect. Agent. Cancer"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1186\/s12985-021-01732-8","article-title":"Prevalence and distribution of human papillomavirus genotypes among women attending gynecology clinics in northern Henan Province of China","volume":"19","author":"Wang","year":"2022","journal-title":"Virol. J."},{"key":"B47","doi-asserted-by":"publisher","first-page":"481","DOI":"10.1097\/cco.0000000000000663","article-title":"Advances in immunotherapy for cervical cancer","volume":"32","author":"Wendel Naumann","year":"2020","journal-title":"Curr. Opin. Oncol."},{"key":"B48","doi-asserted-by":"publisher","DOI":"10.3390\/v11040369","article-title":"Manipulation of epithelial differentiation by HPV oncoproteins","volume":"11","author":"White","year":"2019","journal-title":"Viruses"},{"key":"B49","doi-asserted-by":"publisher","first-page":"220","DOI":"10.1002\/path.1966","article-title":"Increased gene copy numbers at chromosome 20q are frequent in both squamous cell carcinomas and adenocarcinomas of the cervix","volume":"209","author":"Wilting","year":"2006","journal-title":"J. Pathol."},{"key":"B50","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1038\/s41598-017-00136-5","article-title":"Usp7-dependent histone H3 deubiquitylation regulates maintenance of DNA methylation","volume":"7","author":"Yamaguchi","year":"2017","journal-title":"Sci. Rep."},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.3390\/biom8040165","article-title":"RNA Sequencing Provides Insights into the Regulation of Solanesol Biosynthesis in Nicotiana tabacum Induced by Moderately High Temperature","volume":"8","author":"Yan","year":"2018","journal-title":"Biomolecules"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1128\/aac.01513-21","article-title":"Protamine sulfate is a potent inhibitor of human papillomavirus infection in vitro and in vivo","volume":"66","author":"Young","year":"2022","journal-title":"Antimicrob. Agents Chemother."},{"key":"B53","doi-asserted-by":"publisher","first-page":"505","DOI":"10.7150\/ijbs.18834","article-title":"Targeting HSP90-HDAC6 regulating network implicates precision treatment of breast cancer","volume":"13","author":"Yu","year":"2017","journal-title":"Int. J. Biol. Sci."},{"key":"B54","doi-asserted-by":"publisher","first-page":"809","DOI":"10.1089\/hum.2010.071","article-title":"Control of cervicovaginal HPV-16 E7-expressing tumors by the combination of therapeutic HPV vaccination and vascular disrupting agents","volume":"22","author":"Zeng","year":"2011","journal-title":"Hum. Gene Ther."},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2021.677867","article-title":"Novel affibody molecules targeting the HPV16 E6 oncoprotein inhibited the proliferation of cervical cancer cells","volume":"9","author":"Zhu","year":"2021","journal-title":"Front. Cell Dev. Biol."},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21249533","article-title":"p53 is regulated in a biphasic manner in hypoxic human papillomavirus type 16 (HPV16)-positive cervical cancer cells","volume":"21","author":"Zhuang","year":"2020","journal-title":"Int. J. Mol. Sci."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2024.1459572\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,1]],"date-time":"2024-10-01T05:10:37Z","timestamp":1727759437000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2024.1459572\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,1]]},"references-count":56,"alternative-id":["10.3389\/fcimb.2024.1459572"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2024.1459572","relation":{},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,1]]},"article-number":"1459572"}}