{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T23:58:35Z","timestamp":1776211115917,"version":"3.50.1"},"reference-count":26,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,6,18]],"date-time":"2021-06-18T00:00:00Z","timestamp":1623974400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003407","name":"Ministero dell\u2019Istruzione, dell\u2019Universit\u00e0 e della Ricerca","doi-asserted-by":"publisher","award":["PRIN 2017-Grant Number 20174TB8KW"],"award-info":[{"award-number":["PRIN 2017-Grant Number 20174TB8KW"]}],"id":[{"id":"10.13039\/501100003407","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Infection with human papillomavirus type 16 (HPV16) is one of the major risk factors for the development of cervical cancer. Our previous studies have demonstrated the involvement of the early oncoprotein E5 of HPV16 (16E5) in the altered isoform switch of fibroblast growth factor receptor 2 (FGFR2) and the consequent expression in human keratinocytes of the mesenchymal FGFR2c isoform, whose aberrant signaling leads to EMT, invasiveness, and dysregulated differentiation. Here, we aimed to establish the possible direct link between these pathological features or the appearance of FGFR2c and the expression of 16E5 in low-grade squamous intraepithelial lesions (LSILs). Molecular analysis showed that the FGFR2c expression displayed a statistically significant positive correlation with that of the viral oncoprotein, whereas the expression values of the epithelial FGR2b variant, as well as those of the differentiation markers keratin 10 (K10), loricrin (LOR) and involucrin (INV), were inversely linked to the 16E5 expression. In contrast, the expression of EMT-related transcription factors Snail1 and ZEB1 overlapped with that of 16E5, becoming a statistically significant positive correlation in the case of Snail2. Parallel analysis performed in human cervical LSIL-derived W12 cells, containing episomal HPV16, revealed that the depletion of 16E5 by siRNA was able to counteract these molecular events, proving to represent an effective strategy to identify the specific role of this viral oncoprotein in determining LSIL oncogenic and more aggressive profiles. Overall, coupling in vitro approaches to the molecular transcript analysis in ectocervical early lesions could significantly contribute to the characterization of specific gene expression profiles prognostic for those LSILs with a greater probability of direct neoplastic progression.<\/jats:p>","DOI":"10.3390\/ijms22126534","type":"journal-article","created":{"date-parts":[[2021,6,18]],"date-time":"2021-06-18T04:10:47Z","timestamp":1623989447000},"page":"6534","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Expression of the E5 Oncoprotein of HPV16 Impacts on the Molecular Profiles of EMT-Related and Differentiation Genes in Ectocervical Low-Grade Lesions"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5039-0834","authenticated-orcid":false,"given":"Danilo","family":"Ranieri","sequence":"first","affiliation":[{"name":"Department of Clinical and Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Deborah","family":"French","sequence":"additional","affiliation":[{"name":"Department of Clinical and Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy"},{"name":"S. Andrea University Hospital, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9501-7638","authenticated-orcid":false,"given":"Salvatore","family":"Raffa","sequence":"additional","affiliation":[{"name":"Department of Clinical and Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy"},{"name":"S. Andrea University Hospital, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4182-1633","authenticated-orcid":false,"given":"Luisa","family":"Guttieri","sequence":"additional","affiliation":[{"name":"Department of Clinical and Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Rosaria","family":"Torrisi","sequence":"additional","affiliation":[{"name":"Department of Clinical and Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy"},{"name":"S. Andrea University Hospital, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Belleudi","sequence":"additional","affiliation":[{"name":"Department of Clinical and Molecular Medicine, Sapienza University of Rome, 00161 Rome, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,6,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1016\/S0140-6736(13)60022-7","article-title":"Human papillomavirus and cervical cancer","volume":"382","author":"Crosbie","year":"2013","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"F55","DOI":"10.1016\/j.vaccine.2012.06.083","article-title":"The biology and life-cycle of human papillomaviruses","volume":"30","author":"Doorbar","year":"2012","journal-title":"Vaccine"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"4081","DOI":"10.1158\/0008-5472.CAN-09-3335","article-title":"In vitro progression of human papillomavirus 16 episome-associated cervical neoplasia displays fundamental similarities to integrant-associated carcinogenesis","volume":"70","author":"Gray","year":"2010","journal-title":"Cancer Res."},{"key":"ref_4","first-page":"3423","article-title":"Cervical cancer stem cells and other leading factors associated with cervical cancer development","volume":"18","author":"Olmos","year":"2019","journal-title":"Oncol. Lett."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2924","DOI":"10.1158\/0008-5472.CAN-09-3436","article-title":"A role for HPV16 E5 in cervical carcinogenesis","volume":"70","author":"Maufort","year":"2010","journal-title":"Cancer Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1080\/21645515.2017.1264777","article-title":"HPV 16 E5 oncoprotein is expressed in early stage carcinogenesis and can be a target of immunotherapy","volume":"13","author":"Paolini","year":"2017","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_7","doi-asserted-by":"crossref","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":"ref_8","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1080\/19336918.2015.1042197","article-title":"E5 and E6\/E7 of high-risk HPVs cooperate to enhance cancer progression through EMT initiation","volume":"9","year":"2015","journal-title":"Cell Adh Migr."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1186\/s13046-017-0541-1","article-title":"hrHPV E5 oncoprotein: Immune evasion and related immunotherapies","volume":"36","author":"Venuti","year":"2017","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"192","DOI":"10.18632\/oncotarget.803","article-title":"HPV16 E5 and KGFR\/FGFR2b interplay in differentiating epithelial cells","volume":"4","author":"Purpura","year":"2013","journal-title":"Oncotarget"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1038\/s41419-018-0509-x","article-title":"Role of FGFR2b expression and signaling in keratinocyte differentiation: Sequential involvement of PKC\u03b4 and PKC\u03b1","volume":"9","author":"Rosato","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1002\/ijc.29373","article-title":"HPV16 E5 expression induces switching from FGFR2b to FGFR2c and epithelial-mesenchymal transition","volume":"137","author":"Ranieri","year":"2015","journal-title":"Int. J. Cancer"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5440","DOI":"10.18632\/oncotarget.6706","article-title":"Expression of the FGFR2 mesenchymal splicing variant in epithelial cells drives epithelial-mesenchymal transition","volume":"7","author":"Ranieri","year":"2016","journal-title":"Oncotarget"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1002\/mc.22754","article-title":"Expression of the FGFR2c mesenchymal splicing variant in human keratinocytes inhibits differentiation and promotes invasion","volume":"57","author":"Ranieri","year":"2018","journal-title":"Mol. Carcinog."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.virol.2018.03.021","article-title":"Human papillomavirus type 16 E5-mediated upregulation of Met in human keratinocytes","volume":"519","author":"Scott","year":"2018","journal-title":"Virology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"103581","DOI":"10.18632\/oncotarget.21658","article-title":"Human papillomavirus type 18 E5 oncogene supports cell cycle progression and impairs epithelial differentiation by modulating growth factor receptor signalling during the virus life cycle","volume":"8","author":"Wasson","year":"2017","journal-title":"Oncotarget"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1002\/ijc.2910430422","article-title":"Properties of a non-tumorigenic human cervical keratinocyte cell line","volume":"43","author":"Stanley","year":"1989","journal-title":"Int. J. Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1186\/1476-4598-12-38","article-title":"Expression of HPV16 E5 down-modulates the TGFbeta signaling pathway","volume":"12","author":"French","year":"2013","journal-title":"Mol. Cancer"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.jcv.2011.06.007","article-title":"In vivo HPV 16 E5 mRNA: Expression pattern in patients with squamous intra-epithelial lesions of the cervix","volume":"52","author":"Lorenzon","year":"2011","journal-title":"J. Clin. Virol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/j.virusres.2007.09.006","article-title":"The alternative splicing factor hnRNP A1 is up-regulated during virus-infected epithelial cell differentiation and binds the human papillomavirus type 16 late regulatory element","volume":"131","author":"Cheunim","year":"2008","journal-title":"Virus Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1001\/jama.287.16.2114","article-title":"The 2001 Bethesda System: Terminology for reporting results of cervical cytology","volume":"287","author":"Solomon","year":"2002","journal-title":"JAMA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1083\/jcb.106.3.761","article-title":"Normal keratinization in a spontaneously immortalized aneuploid human kera- tinocyte cell line","volume":"106","author":"Boukamp","year":"1988","journal-title":"J. Cell Biol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1359","DOI":"10.1158\/0008-5472.CAN-03-3214","article-title":"Acquisition of high-level chromosomal instability is associated with integration of human papillomavirus type 16 in cervical keratinocytes","volume":"64","author":"Pett","year":"2004","journal-title":"Cancer Res."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Persechino, F., Ranieri, D., Guttieri, L., Nanni, M., Torrisi, M.R., and Belleudi, F. (2021). Expression Profile of Fibroblast Growth Factor Receptors, Keratinocyte Differentiation Markers, and Epithelial Mesenchymal Transition-Related Genes in Actinic Keratosis: A Possible Predictive Factor for Malignant Progression?. Biology, 10.","DOI":"10.3390\/biology10040331"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1093\/carcin\/bgn236","article-title":"Human papillomavirus E5 protein induces expression of the EP4 subtype of prostaglandin E2 receptor in cyclic AMP response element-dependent pathways in cervical cancer cells","volume":"30","author":"Oh","year":"2009","journal-title":"Carcinogenesis"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, I., Rozen, S., and Madden, T.L. (2012). Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction. BMC Bioinform., 13.","DOI":"10.1186\/1471-2105-13-134"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/12\/6534\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:18:09Z","timestamp":1760163489000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/12\/6534"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,6,18]]},"references-count":26,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,6]]}},"alternative-id":["ijms22126534"],"URL":"https:\/\/doi.org\/10.3390\/ijms22126534","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,6,18]]}}}