Recombinant adenoviruses expressing HPV16/18 E7 - upregulate the HDAC6 and DNMT3B genes in C33A cells

Sep 21, 2026

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High-risk human papillomavirus (HPV) drives cervical and other anogenital/oral cancers through oncogenic proteins E6 and E7, whose host regulatory mechanisms remain incompletely defined. Recombinant adenoviruses were constructed to express HPV16/18 E6/E7, delivered into C33A cells for sustained expression, and analyzed using RNA-Seq to map HPV infection-associated transcriptome changes. Results showed E6/E7 overexpression upregulated HDAC6 and DNMT3B and correlated with key cancer-related pathways, supported by qRT-PCR and Western blot. Findings suggest therapeutic target potential and a basis for further E6/E7 oncogenic mechanism studies.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages15

Reading Time38 min

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Introduction

Methods

Results

Conclusion

TYPE Original Research PUBLISHED 01 October 2024 DOI 10.3389/fcimb.2024.1459572 OPEN ACCESS EDITED BY Shih-Heng Chen, National Institute of Environmental Health Sciences (NIH), United States REVIEWED BY Nan Zheng, Nanjing University, China Irene Lo Cigno, University of Eastern Piedmont, Italy *CORRESPONDENCE Jun Wang [wj202fck@163.com](wj202fck@163.com)[ ](wj202fck@163.com)Peng Wang [hellenhello@sina.com](hellenhello@sina.com) RECEIVED 04 July 2024 ACCEPTED 05 September 2024 PUBLISHED 01 October 2024 CITATION Shao Y, Shah PT, Su Q, Li S, Huang F, Wang J, Wang P and Wu C (2024) Recombinant adenoviruses expressing HPV16/18 E7 upregulate the HDAC6 and DNMT3B genes in C33A cells. Front. Cell. Infect. Microbiol. 14:1459572 . doi: 10.3389/fcimb.2024.1459572 COPYRIGHT © 2024 Shao, Shah, Su, Li, Huang, Wang, Wang and Wu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) . The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Recombinant adenoviruses expressing HPV16/18 E7 upregulate the HDAC6 and DNMT3B genes in C33A cells Yunting Shao 1, Pir Tariq Shah 1, Qisheng Su 1, Shanhu Li 2, Fang Huang 2, Jun Wang 3*, Peng Wang 2* and Chengjun Wu 1 1 Faculty of Medicine, School of Basic Medical Sciences, Dalian University of Technology, Dalian, China, 2 Department of Cell Engineering, Beijing Institute of Biotechnology, Beijing, China, 3Obstetrics and Gynecology Department, The Second Afﬁliated Hospital of Dalian Medical University Mailing, Dalian, China Objective: 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. Methods: 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 fortranscriptome sequencing to identify differentially expressed genes associated with HPV infection in cervical cancer. Results: 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. Conclusion: 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 Frontiers in Cellular and Infection Microbiology 01 [fronti

FAQs

What was the study objective regarding HPV E6/E7 proteins?

To clarify how the HPV oncogenic proteins E6 and E7 regulate host processes, focusing on downstream gene changes relevant to HPV-related cervical cancer.

How were recombinant adenoviruses prepared and used in the experiments?

Recombinant adenoviral particles were constructed to express HPV16 and HPV18 E6/E7, then used to infect C33A cells to achieve sustained expression of E6/E7 before transcriptome analysis.

Which genes and pathways were identified as upregulated after E6/E7 expression?

RNA-Seq showed upregulation of HDAC6 and DNMT3B, and correlation analyses linked these genes to pathways including DNA replication, tumor proliferation signatures, G2M checkpoint, p53 pathways, and PI3K/AKT/mTOR signaling.

Sep 21, 2026

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