Progressive p53 Expression in the Histological Spectrum of Cervical Neoplasia

Progressive p53 Expression in the Histological Spectrum of Cervical Neoplasia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Progressive p53 Expression in the Histological Spectrum of Cervical Neoplasia Samuel Ayobami FASOGBON, Frederick Olusegun AKINBO, Comfort E. WILLIAMS, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7567894/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 14 You are reading this latest preprint version Abstract Background The development of cervical cancer is significantly influenced by the dysregulation of the tumor suppressor p53. While TP53 mutations are frequently present in HPV-independent malignancies, as evidenced by abnormal immunohistochemistry (IHC) patterns, E6-mediated degradation in HPV-associated illness reduces p53 function. This study examined p53 expression in a Nigerian cohort with cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC) to elucidate its function in lesion progression. Methods Two hundred cervical specimens (CIN I–III and SCC) that were archived as formalin-fixed paraffin-embedded (FFPE) were recovered. For histological confirmation, sections (3 µm) were stained with hematoxylin–eosin (H&E). Then, p53 IHC was performed using the avidin–biotin complex method with microwave antigen retrieval. Proportion and intensity were combined to create a semi-quantitative score (0–6) for the expression. Types: low (1–3), high (4–6), and negative (0). In SPSS v16, associations with age and grade were examined using χ². Results High levels of p53 expression were seen in 60.9% of SCC and 46.6% of CIN. CIN I (20.9%), CIN II (70.2%), CIN III (46.3%), and SCC (60.9%) are the grades. There was a significant correlation (p < 0.05) between expression and lesion severity. Conclusion p53 expression rises with the severity of the dysplasia, indicating that it may be used as an auxiliary biomarker of the development of cervical neoplasia. Combining pattern-based p53 interpretation, p16/Ki-67 dual-stain, and HPV genotyping may improve its diagnostic and prognostic utility. Cervical intraepithelial neoplasia p53 squamous cell carcinoma immunohistochemistry biomarker human papillomavirus tumor suppressor genes Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction A significant public health concern is still cervical cancer, especially in sub-Saharan Africa, where the disease burden is high and screening coverage is poor [1, 2]. Persistent high-risk HPV infection is the main histological factor responsible for the development of cervical intraepithelial neoplasia (CIN) to invasive cancer. Cell-cycle regulation and genomic stability are compromised by the viral oncoproteins E6 and E7, which interfere with the p53 and RB pathways, respectively. Cervical squamous lesions have been reclassified into HPV-associated and HPV-independent pathways in the 5th edition of the WHO classification. Despite being less frequent, HPV-independent squamous carcinomas have worse prognoses and are distinguished by abnormal p53 IHC patterns and frequent TP53 mutations [3]. The necessity to more precisely identify p53 expression in CIN and aggressive cancer is highlighted by this distinction. An effective method for examining the p53 protein status in archival material is immunohistochemistry (IHC). Wild-type (patchy/variable) and aberrant (diffuse overexpression, complete absence/null, or cytoplasmic) are distinguished by pattern-based interpretation [4]. Even though p53 is not included in standard screening algorithms, it could be useful for understanding the biology of diseases and for supplementary diagnostics, particularly in cases where HPV-independent illness is suspected. The importance of other biomarkers is also highlighted by recent research; for instance, Fasogbon & Akinbo [1] found that the expression of the vitamin D receptor (VDR) gradually decreases with increasing CIN grades, supporting the idea that the tumor suppressor pathway is disrupted. This study adds local evidence to the global understanding of cervical tumor suppressor biology by examining the expression profile of p53 across CIN grades and SCC in a Nigerian cohort. Materials and Methods Study design and sample selection: Two hundred archival FFPE cervical tissue blocks diagnosed at Lagos University Teaching Hospital between 2002 and 2016 were the subject of a retrospective, cross-sectional analysis. Invasive SCC and CIN I, CIN II, and CIN III cases were among them. Sufficient tissue, a verified histological diagnosis, and the availability of clinical age data are requirements for inclusion. Blocks with inadequate preservation or unclear diagnoses are excluded. Ethical approval: Granted approval by the Lagos University Teaching Hospital Health Research Ethics Committee (LUTHHREC) on June 18, 2020, under approval number ADM/DCST/HREC/APP/3429. This study complied with the Helsinki Declaration. Histopathological review: Sections of 3 μm were cut for every case. To assess lesion adequacy and confirm the diagnosis, one section was stained with H&E. Immunohistochemistry: IHC for p53 was carried out using the avidin–biotin complex (ABC) method using a mouse monoclonal anti-p53 antibody. Using microwave heating, antigen retrieval was performed in citrate buffer (pH 6.0) [9]. Primary antibody (1:100, 60 min), secondary antibody, HRP conjugate, and DAB chromogen were treated with the sections. Mayer's hematoxylin was used for counterstaining. Both negative controls (primary antibody deleted) and appropriate positive controls (known p53-positive cancer) were included. Scoring: A semi-quantitative assessment of nuclear staining was conducted [13]: The proportions are as follows: 0 = 0%, 1 = 60%. • Intensity is denoted by 0 for none, 1 for weak, 2 for moderate, and 3 for strong. • A + B (0–6) is the final score. The scores were divided into three categories: low (1–3), high (4–6), and negative (0). To increase reproducibility, two assessors separately examined each slide; disagreements were settled by consensus. Statistical analysis: SPSS v16 was used to enter the data. Expression levels were summarized using descriptive statistics. Chi-square was used to look for associations between p53 category and age or histological grade; a p-value of less than 0.05 was deemed statistically significant. Results According to Table 1, high p53 expression was seen in 60.9% of SCC and 46.6% of CIN. SCC (60.9%), CIN I (20.9%), CIN II (70.2%), and CIN III (46.3%) by grade Lesion severity and expression had a strong correlation (p < 0.05). The highest incidence of high expression was found in women between the ages of 41 and 50 (Table 2). Table 1. Expression of p53 in CIN I–III and SCC Case type n tested High expression (%) Low expression (%) p-value CIN I 43 9 (20.9) 34 (79.1) CIN II 47 33 (70.2) 14 (29.8) CIN III 41 19 (46.3) 22 (53.7) SCC 69 42 (60.9) 27 (39.1) <0.05 Table 2. Age distribution and p53 expression in CIN and SCC Age group (years) High expression (%) Low expression (%) Total (%) ≤40 14 (12.1) 19 (16.2) 33 (28.3) 41–50 46 (20.4) 41 (18.1) 87 (38.5) ≥50 43 (19.0) 37 (14.2) 80 (33.2) Total 103 (51.5) 97 (48.5) 200 (100) Figures 1–4 display representative immunohistochemical pictures that indicate the progression and distribution of p53 expression in CIN I–III and SCC. Discussion In line with data from Europe, Asia, and Africa [10–12], this study showed a gradual rise in p53 expression from CIN I through SCC. Our results are consistent with the idea of a molecular continuum where histological advancement is accompanied by a progressive deregulation of tumor suppressor pathways. Biological interpretation There are differences between HPV-associated and HPV-independent pathways in the processes behind p53 disruption. The increase of wild-type protein under stress or the stabilization of mutant protein may be reflected in p53 upregulation in HPV-associated lesions [14,15]. TP53 mutations are frequently found in HPV-independent carcinomas, which can lead to abnormal IHC patterns including diffuse high positivity or total absence [6,7]. Identifying these patterns can be useful for diagnosis, especially when HPV testing comes back negative. Comparison with other biomarkers Biomarker panels work better than individual markers, according to recent studies. The most proven adjuncts for diagnosis and triage are still p16 and ki-67 [8]. In India, Kaur et al. (2025) showed a high correlation between CIN grade and the combined expression of p53 and Ki-67. Concordant overexpression of p16, p53, and Ki-67 was reported in Nigeria by Ogbu et al. (2022), highlighting the significance of these genes in African populations. In line with the p53 trajectory, our earlier research on the vitamin D receptor (VDR) revealed increasing receptor loss across CIN grades [1]. These results collectively imply that cumulative deactivation of several tumor suppressor mechanisms plays a role in cervical carcinogenesis. Clinical implications Although p53 has potential as an auxiliary biomarker in pathology, it is not advised for primary screening. It may help in the following: Clarification of unclear histology instances is one area. Comprehending HPV-independent carcinoma, which has a poorer outlook. Aiding in diagnosis in cases where molecular resources are scarce. Incorporating p53 into pathology workflows in addition to p16/Ki-67 and HPV DNA tests may improve diagnosis accuracy and help guide treatment plans. Strengths and limitations The inclusion of all CIN grades, the comparatively high sample size, and the underrepresented local African data are among its strengths. Retrospective design, lack of HPV type, lack of outcome data, and reliance on semi-quantitative interpretation rather than pattern-based interpretation, which is now advised by WHO/ESMO guidelines, are among the limitations. Future perspectives Future studies should: Use molecular markers and HPV genotyping to more accurately categorize diseases that are not caused by HPV. Use pattern-based p53 scoring to differentiate expression linked to the wild type from that of the mutant. Investigate AI-assisted quantification and digital pathology to lessen observer variability. Perform prospective African cohort studies with clinical follow-up to confirm prognostic significance. Conclusion The molecular continuum of cervical carcinogenesis is shown in this study, which shows that p53 expression rises gradually from CIN to invasive squamous carcinoma. Our results, which come from a Nigerian cohort, add to the scant information available from sub-Saharan Africa and highlight the usefulness of p53 as a secondary biomarker as opposed to a main screening method. Although p53 cannot replace HPV testing or p16/Ki-67 dual-stain, it can aid in histopathological grading, especially in lesions that are difficult to diagnose or HPV-independent. To confirm its predictive value and improve risk classification in a variety of groups, future research involving HPV genotyping, pattern-based p53 interpretation, and other biomarkers is required. Declarations Ethics approval and consent to participate Granted approval by the Lagos University Teaching Hospital Health Research Ethics Committee (LUTHHREC) on June 18, 2020, under approval number ADM/DCST/HREC/APP/3429. According to national regulations, the Ethics Committee exempted the informed consent requirement because this was a retrospective study employing anonymized, archived formalin-fixed paraffin-embedded (FFPE) tissue blocks. Consent for publication Not applicable. Availability of data and materials This published article already contains the datasets created and examined during the current investigation. The Corresponding author can provide more supporting information upon reasonable request. Competing interests The author declares no competing interests. Acknowledgements The authors would like to express their gratitude to Mr. Osiagwu Daniel of the University of Lagos College of Medicine and Dr. Jonathan U. Madukwe of the Cellular Pathology Specialty at the National Hospital Abuja for their technical support of the immunohistochemistry laboratory study. We also sincerely thank Prof. A.A.F. Banjo for his important contribution to tissue diagnostics. Funding This study did not get any special funding. Authors’ contributions Samuel Ayobami Fasogbon (SAF): Developed and planned the research, carried out immunohistochemistry, analyzed data, interpreted findings, and wrote the manuscript. Frederick Olusegun Akinbo (FOA): Provided supervision, contributed to study design, guided interpretation of results, and critically revised the manuscript for important intellectual content. Comfort E. Williams (CEW): Assisted in histopathological review and confirmation of tissue diagnoses. Charles Egede Ugwu (CEU): Contributed to laboratory support and immunohistochemistry slide preparation. Bob-Manuel Chinonso Osuji (BCO): Performed statistical analysis, contributed to data interpretation, and supported results validation. Peter O. Obami (POO): Assisted in manuscript writing, contextual discussion, and final critical review. All authors read and approved the final version of the manuscript. References Fasogbon SA, Akinbo FO. Vitamin D receptor protagonist in cervical neoplasia grades. Egypt J Basic Appl Sci . 2025;12(1):1–10. doi:10.1080/2314808X.2024.2434992. Perkins RB, Guido RS, Castle PE, et al. 2019 ASCCP risk-based management consensus guidelines. J Low Genit Tract Dis . 2020;24(2):102–131. doi:10.1097/LGT.0000000000000525. Höhn AK, Brambs CE, Dannecker C, Diebold J. The 2020 WHO classification of female genital tumors. Geburtsh Frauenheilkd . 2021;81(10):1145–1153. doi:10.1055/a-1545-4279. Tessier-Cloutier B, Kortekaas KE, Thompson EF, et al. Major p53 IHC patterns correlate with TP53 mutation status. Mod Pathol . 2020;33(3):574–583. doi:10.1038/s41379-019-0409-7. Thompson EF, Mittal K, Soslow RA, et al. p53 IHC patterns in HPV-related neoplasms. Mod Pathol . 2020;33(9):1653–1663. doi:10.1038/s41379-020-0528-5. Banister CE, Liu C, Pirisi L, Creek KE, Buckhaults PJ. Identification of HPV-independent cervical cancers. Oncotarget . 2017;8(7):13375–13386. doi:10.18632/oncotarget.14562. Lee JE, Park JS, Nam EJ, et al. HPV-negative cervical cancers: molecular features and TP53 mutations. BMB Rep . 2022;55(9):442–450. doi:10.5483/BMBRep.2022.55.9.135. Clarke MA, Cheung LC, Castle PE, et al. Recommendations for use of p16/Ki-67 dual stain. J Low Genit Tract Dis . 2024;28(2):e1–e12. doi:10.1097/LGT.0000000000000784. Shi SR, Key ME, Kalra KL. Antigen retrieval in FFPE tissues. J Histochem Cytochem . 1991;39(6):741–748. doi:10.1177/39.6.1709656. Giarnieri E, Zanconati F, Floreani A, et al. p53 overexpression in cervical lesions. J Clin Pathol . 2000;53(6):445–449. doi:10.1136/jcp.53.6.445. Cheah PL, Looi LM. p53 expression in cervical squamous lesions. Pathology . 2002;34(4):326–331. doi:10.1080/00313020220131392. Grace VM, Meleth S, Rajeshkumar NV, et al. p53 alterations in CIN and carcinoma. Int J Gynecol Pathol . 2003;22(4):383–388. doi:10.1097/00004347-200310000-00009. Klein F, et al. Immunohistochemical scoring systems in oncology. Histopathology . 1999;35(3):232–238. doi:10.1046/j.1365-2559.1999.00643.x. Mantovani F, Collavin L, Del Sal G. Mutant p53 as a guardian turned killer. Nat Rev Cancer . 2019;19(10):651–667. doi:10.1038/s41568-019-0177-4. Vousden KH, Prives C. Blinded by the Light: The Growing Complexity of p53. Cell . 2009;137(3):413–431. doi:10.1016/j.cell.2009.04.037. Kaur M, et al. Study of Ki-67 and p53 expression in cervical neoplasia. Healthcare Bulletin . 2025;15(5):639–645. Ogbu CC, et al. Correlation of p16, p53, Ki-67 in noninvasive cervical lesions. J Adv Surg Clin Res . 2022;12(3):25–32. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 30 Oct, 2025 Reviews received at journal 29 Oct, 2025 Reviewers agreed at journal 21 Oct, 2025 Reviews received at journal 18 Oct, 2025 Reviewers agreed at journal 18 Oct, 2025 Reviews received at journal 13 Oct, 2025 Reviewers agreed at journal 09 Oct, 2025 Reviewers agreed at journal 08 Oct, 2025 Reviewers agreed at journal 08 Oct, 2025 Reviewers invited by journal 06 Oct, 2025 Editor assigned by journal 18 Sep, 2025 Editor invited by journal 10 Sep, 2025 Submission checks completed at journal 09 Sep, 2025 First submitted to journal 09 Sep, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7567894","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":530690220,"identity":"03df6fe8-1f10-4b82-a523-81b5bea08a00","order_by":0,"name":"Samuel Ayobami FASOGBON","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBElEQVRIiWNgGAWjYBAC+QYGhg8MDAeYDUA8IIuBjZ2AFoMDDIwzgFrYgVoYG2eAtDAT0sIA0cIPYjTzgIQIamE/ndjw488daXP29uePbX5tk+djZmD88DEHj196cjc29rY9M7bsOWPYnNt327CNmYFZcuY2PNYcyN3+gLfhcLLBjRzG5tye24xALWzMvPi0nH+7sfHPn8P1G+4/f9hs2XPbnrCWG7kbm3nYDjMb3GAwbGb4cTuRoBaDG283Nsu2HWa27MkxnNnbcDu5jZmxGa9f5PuB3n/z5zCzOfvxBx9+/LltO7+9+eCHj/gchgIY28BkA7HqQeAPKYpHwSgYBaNgpAAAeWhcji/EHUAAAAAASUVORK5CYII=","orcid":"","institution":"Medical Laboratory Science Council of Nigeria","correspondingAuthor":true,"prefix":"","firstName":"Samuel","middleName":"Ayobami","lastName":"FASOGBON","suffix":""},{"id":530690221,"identity":"384a7b55-334b-4593-962a-50f6a1210018","order_by":1,"name":"Frederick Olusegun AKINBO","email":"","orcid":"","institution":"University of Benin","correspondingAuthor":false,"prefix":"","firstName":"Frederick","middleName":"Olusegun","lastName":"AKINBO","suffix":""},{"id":530690222,"identity":"9a6e5fdf-fcd8-4484-8208-ba7e2887e269","order_by":2,"name":"Comfort E. WILLIAMS","email":"","orcid":"","institution":"New York Medical College","correspondingAuthor":false,"prefix":"","firstName":"Comfort","middleName":"E.","lastName":"WILLIAMS","suffix":""},{"id":530690223,"identity":"037945c9-eb0b-4605-b3d0-c3323fe5e3a9","order_by":3,"name":"Charles Egede UGWU","email":"","orcid":"","institution":"Washington State University","correspondingAuthor":false,"prefix":"","firstName":"Charles","middleName":"Egede","lastName":"UGWU","suffix":""},{"id":530690224,"identity":"701874a0-64cc-4595-a43d-8ec26dfe5793","order_by":4,"name":"Bob-Manuel Chinonso OSUJI","email":"","orcid":"","institution":"Teesside University","correspondingAuthor":false,"prefix":"","firstName":"Bob-Manuel","middleName":"Chinonso","lastName":"OSUJI","suffix":""},{"id":530690225,"identity":"2ef54273-02ea-4a22-a542-69a680ba23f4","order_by":5,"name":"Peter O. OBAMI","email":"","orcid":"","institution":"Barking, Havering And Redbridge University Hospitals NHS Trust","correspondingAuthor":false,"prefix":"","firstName":"Peter","middleName":"O.","lastName":"OBAMI","suffix":""}],"badges":[],"createdAt":"2025-09-08 23:53:03","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7567894/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7567894/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":93741432,"identity":"9ecc5fa2-618d-4a10-8538-0dc24bdd1524","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":322357,"visible":true,"origin":"","legend":"","description":"","filename":"p53cervicalneoplasiamanuscript1.docx","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/c944b7bbc30518bbd0297fb1.docx"},{"id":93741941,"identity":"4b8c7e8b-3f7d-46e5-a995-b9970d27bd1d","added_by":"auto","created_at":"2025-10-17 05:27:35","extension":"json","order_by":1,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":8560,"visible":true,"origin":"","legend":"","description":"","filename":"49f93399ab8d49ebb1f5a850628874f0.json","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/6ea1d6274aed24f44742b866.json"},{"id":93741431,"identity":"8f26a0a1-805f-42d5-8f98-75e662d41e59","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"xml","order_by":2,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":44072,"visible":true,"origin":"","legend":"","description":"","filename":"49f93399ab8d49ebb1f5a850628874f01enriched.xml","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/0a4718fad17d98b7e83a7331.xml"},{"id":93741434,"identity":"3c6706d2-41a0-431a-9105-43ba3971b289","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"jpeg","order_by":3,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1089173,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/133abfe0ae2582bb1ae30056.jpeg"},{"id":93741441,"identity":"5324ce35-0fe7-4363-9d25-0ce23c31a4be","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"jpeg","order_by":4,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1203983,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/5c2882e94cfe5cf7c63fa484.jpeg"},{"id":93741437,"identity":"9c284a04-56b7-43a3-941d-6c0be599412e","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"jpeg","order_by":5,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1285252,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage3.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/7ac64dd52e3c09f0213089fb.jpeg"},{"id":93741447,"identity":"bc49bf10-8d65-4bfb-a49e-3ed9c252b286","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"jpeg","order_by":6,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":1033165,"visible":true,"origin":"","legend":"","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/666b69d32b508a0351ee18ed.jpeg"},{"id":93741448,"identity":"83256f4b-e04f-416e-8a9d-f3e6d13dbfc1","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"png","order_by":7,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":365096,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/53ed7fbadf8cf15d26dd6e99.png"},{"id":93741942,"identity":"b674099e-76e1-4d60-8f25-5836e39520ce","added_by":"auto","created_at":"2025-10-17 05:27:35","extension":"png","order_by":8,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":445022,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/f612963f10ffe5891efcb53e.png"},{"id":93742220,"identity":"5d02c02c-4f13-4366-a889-b374e7376152","added_by":"auto","created_at":"2025-10-17 05:35:37","extension":"png","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":459010,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/7ebae682052253dbc1539dcd.png"},{"id":93741443,"identity":"7389edf2-4dd6-484c-b527-0b7c27c053ab","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"png","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":395018,"visible":true,"origin":"","legend":"","description":"","filename":"Onlinefloatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/0ce88a5d5f7e015705bdb638.png"},{"id":93741444,"identity":"fa0bc241-4894-46bc-a384-0be0854ed64e","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"xml","order_by":11,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":39226,"visible":true,"origin":"","legend":"","description":"","filename":"49f93399ab8d49ebb1f5a850628874f01structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/399dcb5665c0bcbc22628aec.xml"},{"id":93741446,"identity":"325754b9-adce-47cb-857b-cf552eb3e2e1","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"html","order_by":12,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":49880,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/461e3ea880b80b6f2aa12c1e.html"},{"id":93741439,"identity":"acfdaf38-5b37-4056-a7f2-266e192157b7","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":955197,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemistry of mild expression of p53 in CIN I showing dysplastic cells confined to the lower third of the epithelium with brown nuclear staining (x400).\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/5eabf32521272d31a65780e9.png"},{"id":93741445,"identity":"75e3aca0-e57a-439c-af8b-b986ae3d4f0c","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1080615,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemistry of moderate expression of p53 in CIN II showing atypical cells in the lower two-thirds of the epithelium with brown nuclear staining (x400).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/f24c769789e29285eba120d5.png"},{"id":93741433,"identity":"fa26d143-d3d7-487f-82d5-f8683d360525","added_by":"auto","created_at":"2025-10-17 05:19:35","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1122040,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemistry of strong expression of p53 in CIN III showing full-thickness dysplasia with loss of polarity and strong brown nuclear staining (x400).\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/a5e4f11e54ed76b8fb340e52.png"},{"id":93741940,"identity":"2963e2be-2220-4d96-a3a6-29d518d27af3","added_by":"auto","created_at":"2025-10-17 05:27:35","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":898415,"visible":true,"origin":"","legend":"\u003cp\u003eImmunohistochemistry of strong expression of p53 in squamous cell carcinoma showing nests of malignant squamous cells infiltrating the stroma with diffuse brown nuclear staining (x400).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/e70dba74eb7535b9f3ce1ac0.png"},{"id":93742739,"identity":"52ca0e90-fb06-4d03-914f-55c5ebe4db64","added_by":"auto","created_at":"2025-10-17 05:43:40","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6450799,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7567894/v1/e7d547dc-8d2a-4fdf-b36e-f5c904d77833.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Progressive p53 Expression in the Histological Spectrum of Cervical Neoplasia","fulltext":[{"header":"Introduction","content":"\u003cp\u003eA significant public health concern is still cervical cancer, especially in sub-Saharan Africa, where the disease burden is high and screening coverage is poor [1, 2]. Persistent high-risk HPV infection is the main histological factor responsible for the development of cervical intraepithelial neoplasia (CIN) to invasive cancer. Cell-cycle regulation and genomic stability are compromised by the viral oncoproteins E6 and E7, which interfere with the p53 and RB pathways, respectively.\u003c/p\u003e\u003cp\u003eCervical squamous lesions have been reclassified into HPV-associated and HPV-independent pathways in the 5th edition of the WHO classification. Despite being less frequent, HPV-independent squamous carcinomas have worse prognoses and are distinguished by abnormal p53 IHC patterns and frequent TP53 mutations [3]. The necessity to more precisely identify p53 expression in CIN and aggressive cancer is highlighted by this distinction.\u003c/p\u003e\u003cp\u003eAn effective method for examining the p53 protein status in archival material is immunohistochemistry (IHC). Wild-type (patchy/variable) and aberrant (diffuse overexpression, complete absence/null, or cytoplasmic) are distinguished by pattern-based interpretation [4].\u003c/p\u003e\u003cp\u003eEven though p53 is not included in standard screening algorithms, it could be useful for understanding the biology of diseases and for supplementary diagnostics, particularly in cases where HPV-independent illness is suspected. The importance of other biomarkers is also highlighted by recent research; for instance, Fasogbon \u0026amp; Akinbo [1] found that the expression of the vitamin D receptor (VDR) gradually decreases with increasing CIN grades, supporting the idea that the tumor suppressor pathway is disrupted.\u003c/p\u003e\u003cp\u003eThis study adds local evidence to the global understanding of cervical tumor suppressor biology by examining the expression profile of p53 across CIN grades and SCC in a Nigerian cohort.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eStudy design and sample selection:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTwo hundred archival FFPE cervical tissue blocks diagnosed at Lagos University Teaching Hospital between 2002 and 2016 were the subject of a retrospective, cross-sectional analysis. Invasive SCC and CIN I, CIN II, and CIN III cases were among them. Sufficient tissue, a verified histological diagnosis, and the availability of clinical age data are requirements for inclusion. Blocks with inadequate preservation or unclear diagnoses are excluded.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGranted approval by the Lagos University Teaching Hospital Health Research Ethics Committee (LUTHHREC) on June 18, 2020, under approval number ADM/DCST/HREC/APP/3429. This study complied with the Helsinki Declaration.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHistopathological review:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSections of 3 \u0026mu;m were cut for every case. To assess lesion adequacy and confirm the diagnosis, one section was stained with H\u0026amp;E.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eImmunohistochemistry:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIHC for p53 was carried out using the avidin\u0026ndash;biotin complex (ABC) method using a mouse monoclonal anti-p53 antibody. Using microwave heating, antigen retrieval was performed in citrate buffer (pH 6.0) [9]. Primary antibody (1:100, 60 min), secondary antibody, HRP conjugate, and DAB chromogen were treated with the sections. Mayer\u0026apos;s hematoxylin was used for counterstaining. Both negative controls (primary antibody deleted) and appropriate positive controls (known p53-positive cancer) were included.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eScoring:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA semi-quantitative assessment of nuclear staining was conducted [13]:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe proportions are as follows: 0 = 0%, 1 = \u0026lt;30%, 2 = 30%\u0026ndash;60%, and 3 = \u0026gt;60%.\u003c/p\u003e\n\u003cp\u003e\u0026bull; Intensity is denoted by 0 for none, 1 for weak, 2 for moderate, and 3 for strong.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026bull; A + B (0\u0026ndash;6) is the final score.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe scores were divided into three categories: low (1\u0026ndash;3), high (4\u0026ndash;6), and negative (0). To increase reproducibility, two assessors separately examined each slide; disagreements were settled by consensus.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis:\u003c/strong\u003e\u003cbr\u003e\u0026nbsp;SPSS v16 was used to enter the data. Expression levels were summarized using descriptive statistics. Chi-square was used to look for associations between p53 category and age or histological grade; a p-value of less than 0.05 was deemed statistically significant.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eAccording to Table 1, high p53 expression was seen in 60.9% of SCC and 46.6% of CIN.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSCC (60.9%), CIN I (20.9%), CIN II (70.2%), and CIN III (46.3%) by grade Lesion severity and expression had a strong correlation (p \u0026lt; 0.05). The highest incidence of high expression was found in women between the ages of 41 and 50 (Table 2).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1. Expression of p53 in CIN I\u0026ndash;III and SCC\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCase type\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003en tested\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHigh expression (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLow expression (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003ep-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCIN I\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e9 (20.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e34 (79.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCIN II\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e33 (70.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e14 (29.8)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eCIN III\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e19 (46.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e22 (53.7)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eSCC\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e42 (60.9)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e27 (39.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;0.05\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. Age distribution and p53 expression in CIN and SCC\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eAge group (years)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 166px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eHigh expression (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 156px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eLow expression (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 121px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal (%)\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026le;40\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 166px;\"\u003e\n \u003cp\u003e14 (12.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 156px;\"\u003e\n \u003cp\u003e19 (16.2)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 121px;\"\u003e\n \u003cp\u003e33 (28.3)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e41\u0026ndash;50\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 166px;\"\u003e\n \u003cp\u003e46 (20.4)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 156px;\"\u003e\n \u003cp\u003e41 (18.1)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 121px;\"\u003e\n \u003cp\u003e87 (38.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026ge;50\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 166px;\"\u003e\n \u003cp\u003e43 (19.0)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 156px;\"\u003e\n \u003cp\u003e37 (14.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 121px;\"\u003e\n \u003cp\u003e80 (33.2)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 148px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eTotal\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 166px;\"\u003e\n \u003cp\u003e103 (51.5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 156px;\"\u003e\n \u003cp\u003e97 (48.5)\u003c/p\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 121px;\"\u003e\n \u003cp\u003e200 (100)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFigures 1\u0026ndash;4 display representative immunohistochemical pictures that indicate the progression and distribution of p53 expression in CIN I\u0026ndash;III and SCC.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn line with data from Europe, Asia, and Africa [10\u0026ndash;12], this study showed a gradual rise in p53 expression from CIN I through SCC. Our results are consistent with the idea of a molecular continuum where histological advancement is accompanied by a progressive deregulation of tumor suppressor pathways.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBiological interpretation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere are differences between HPV-associated and HPV-independent pathways in the processes behind p53 disruption. The increase of wild-type protein under stress or the stabilization of mutant protein may be reflected in p53 upregulation in HPV-associated lesions [14,15]. TP53 mutations are frequently found in HPV-independent carcinomas, which can lead to abnormal IHC patterns including diffuse high positivity or total absence [6,7]. Identifying these patterns can be useful for diagnosis, especially when HPV testing comes back negative.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eComparison with other biomarkers\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBiomarker panels work better than individual markers, according to recent studies. The most proven adjuncts for diagnosis and triage are still p16 and ki-67 [8]. In India, Kaur et al. (2025) showed a high correlation between CIN grade and the combined expression of p53 and Ki-67. Concordant overexpression of p16, p53, and Ki-67 was reported in Nigeria by Ogbu et al. (2022), highlighting the significance of these genes in African populations.\u0026nbsp;\u003cbr\u003e\u0026nbsp;In line with the p53 trajectory, our earlier research on the vitamin D receptor (VDR) revealed increasing receptor loss across CIN grades [1]. These results collectively imply that cumulative deactivation of several tumor suppressor mechanisms plays a role in cervical carcinogenesis.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eClinical implications\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAlthough p53 has potential as an auxiliary biomarker in pathology, it is not advised for primary screening. It may help in the following:\u003c/p\u003e\n\u003cul\u003e\n \u003cli\u003eClarification of unclear histology instances is one area.\u003c/li\u003e\n \u003cli\u003eComprehending HPV-independent carcinoma, which has a poorer outlook.\u003c/li\u003e\n \u003cli\u003eAiding in diagnosis in cases where molecular resources are scarce.\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003eIncorporating p53 into pathology workflows in addition to p16/Ki-67 and HPV DNA tests may improve diagnosis accuracy and help guide treatment plans.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStrengths and limitations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe inclusion of all CIN grades, the comparatively high sample size, and the underrepresented local African data are among its strengths. Retrospective design, lack of HPV type, lack of outcome data, and reliance on semi-quantitative interpretation rather than pattern-based interpretation, which is now advised by WHO/ESMO guidelines, are among the limitations.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFuture perspectives\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFuture studies should:\u003c/p\u003e\n\u003col\u003e\n \u003cli\u003eUse molecular markers and HPV genotyping to more accurately categorize diseases that are not caused by HPV.\u003c/li\u003e\n \u003cli\u003eUse pattern-based p53 scoring to differentiate expression linked to the wild type from that of the mutant.\u003c/li\u003e\n \u003cli\u003eInvestigate AI-assisted quantification and digital pathology to lessen observer variability.\u003c/li\u003e\n \u003cli\u003ePerform prospective African cohort studies with clinical follow-up to confirm prognostic significance.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe molecular continuum of cervical carcinogenesis is shown in this study, which shows that p53 expression rises gradually from CIN to invasive squamous carcinoma. Our results, which come from a Nigerian cohort, add to the scant information available from sub-Saharan Africa and highlight the usefulness of p53 as a secondary biomarker as opposed to a main screening method. Although p53 cannot replace HPV testing or p16/Ki-67 dual-stain, it can aid in histopathological grading, especially in lesions that are difficult to diagnose or HPV-independent. To confirm its predictive value and improve risk classification in a variety of groups, future research involving HPV genotyping, pattern-based p53 interpretation, and other biomarkers is required.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGranted approval by the Lagos University Teaching Hospital Health Research Ethics Committee (LUTHHREC) on June 18, 2020, under approval number ADM/DCST/HREC/APP/3429. According to national regulations, the Ethics Committee exempted the informed consent requirement because this was a retrospective study employing anonymized, archived formalin-fixed paraffin-embedded (FFPE) tissue blocks.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis published article already contains the datasets created and examined during the current investigation. The Corresponding author can provide more supporting information upon reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author declares no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to express their gratitude to Mr. Osiagwu Daniel of the University of Lagos College of Medicine and Dr. Jonathan U. Madukwe of the Cellular Pathology Specialty at the National Hospital Abuja for their technical support of the immunohistochemistry laboratory study. We also sincerely thank Prof. A.A.F. Banjo for his important contribution to tissue diagnostics.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study did not get any special funding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cul type=\"disc\"\u003e\n \u003cli\u003eSamuel Ayobami Fasogbon (SAF): Developed\u0026nbsp;and\u0026nbsp;planned\u0026nbsp;the\u0026nbsp;research,\u0026nbsp;carried\u0026nbsp;out\u0026nbsp;immunohistochemistry,\u0026nbsp;analyzed\u0026nbsp;data,\u0026nbsp;interpreted\u0026nbsp;findings,\u0026nbsp;and\u0026nbsp;wrote\u0026nbsp;the\u0026nbsp;manuscript.\u003c/li\u003e\n \u003cli\u003eFrederick Olusegun Akinbo (FOA): Provided supervision, contributed to study design, guided interpretation of results, and critically revised the manuscript for important intellectual content.\u003c/li\u003e\n \u003cli\u003eComfort E. Williams (CEW): Assisted in histopathological review and confirmation of tissue diagnoses.\u003c/li\u003e\n \u003cli\u003eCharles Egede Ugwu (CEU): Contributed to laboratory support and immunohistochemistry slide preparation.\u003c/li\u003e\n \u003cli\u003eBob-Manuel Chinonso Osuji (BCO): Performed statistical analysis, contributed to data interpretation, and supported results validation.\u003c/li\u003e\n \u003cli\u003ePeter O. Obami (POO): Assisted in manuscript writing, contextual discussion, and final critical review.\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003eAll authors read and approved the final version of the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col start=\"1\" type=\"1\"\u003e\n\u003cli\u003eFasogbon SA, Akinbo FO. Vitamin D receptor protagonist in cervical neoplasia grades. \u003cem\u003eEgypt J Basic Appl Sci\u003c/em\u003e. 2025;12(1):1\u0026ndash;10. doi:10.1080/2314808X.2024.2434992.\u003c/li\u003e\n\u003cli\u003ePerkins RB, Guido RS, Castle PE, et al. 2019 ASCCP risk-based management consensus guidelines. \u003cem\u003eJ Low Genit Tract Dis\u003c/em\u003e. 2020;24(2):102\u0026ndash;131. doi:10.1097/LGT.0000000000000525.\u003c/li\u003e\n\u003cli\u003eH\u0026ouml;hn AK, Brambs CE, Dannecker C, Diebold J. The 2020 WHO classification of female genital tumors. \u003cem\u003eGeburtsh Frauenheilkd\u003c/em\u003e. 2021;81(10):1145\u0026ndash;1153. doi:10.1055/a-1545-4279.\u003c/li\u003e\n\u003cli\u003eTessier-Cloutier B, Kortekaas KE, Thompson EF, et al. Major p53 IHC patterns correlate with TP53 mutation status. \u003cem\u003eMod Pathol\u003c/em\u003e. 2020;33(3):574\u0026ndash;583. doi:10.1038/s41379-019-0409-7.\u003c/li\u003e\n\u003cli\u003eThompson EF, Mittal K, Soslow RA, et al. p53 IHC patterns in HPV-related neoplasms. \u003cem\u003eMod Pathol\u003c/em\u003e. 2020;33(9):1653\u0026ndash;1663. doi:10.1038/s41379-020-0528-5.\u003c/li\u003e\n\u003cli\u003eBanister CE, Liu C, Pirisi L, Creek KE, Buckhaults PJ. Identification of HPV-independent cervical cancers. \u003cem\u003eOncotarget\u003c/em\u003e. 2017;8(7):13375\u0026ndash;13386. doi:10.18632/oncotarget.14562.\u003c/li\u003e\n\u003cli\u003eLee JE, Park JS, Nam EJ, et al. HPV-negative cervical cancers: molecular features and TP53 mutations. \u003cem\u003eBMB Rep\u003c/em\u003e. 2022;55(9):442\u0026ndash;450. doi:10.5483/BMBRep.2022.55.9.135.\u003c/li\u003e\n\u003cli\u003eClarke MA, Cheung LC, Castle PE, et al. Recommendations for use of p16/Ki-67 dual stain. \u003cem\u003eJ Low Genit Tract Dis\u003c/em\u003e. 2024;28(2):e1\u0026ndash;e12. doi:10.1097/LGT.0000000000000784.\u003c/li\u003e\n\u003cli\u003eShi SR, Key ME, Kalra KL. Antigen retrieval in FFPE tissues. \u003cem\u003eJ Histochem Cytochem\u003c/em\u003e. 1991;39(6):741\u0026ndash;748. doi:10.1177/39.6.1709656.\u003c/li\u003e\n\u003cli\u003eGiarnieri E, Zanconati F, Floreani A, et al. p53 overexpression in cervical lesions. \u003cem\u003eJ Clin Pathol\u003c/em\u003e. 2000;53(6):445\u0026ndash;449. doi:10.1136/jcp.53.6.445.\u003c/li\u003e\n\u003cli\u003eCheah PL, Looi LM. p53 expression in cervical squamous lesions. \u003cem\u003ePathology\u003c/em\u003e. 2002;34(4):326\u0026ndash;331. doi:10.1080/00313020220131392.\u003c/li\u003e\n\u003cli\u003eGrace VM, Meleth S, Rajeshkumar NV, et al. p53 alterations in CIN and carcinoma. \u003cem\u003eInt J Gynecol Pathol\u003c/em\u003e. 2003;22(4):383\u0026ndash;388. doi:10.1097/00004347-200310000-00009.\u003c/li\u003e\n\u003cli\u003eKlein F, et al. Immunohistochemical scoring systems in oncology. \u003cem\u003eHistopathology\u003c/em\u003e. 1999;35(3):232\u0026ndash;238. doi:10.1046/j.1365-2559.1999.00643.x.\u003c/li\u003e\n\u003cli\u003eMantovani F, Collavin L, Del Sal G. Mutant p53 as a guardian turned killer. \u003cem\u003eNat Rev Cancer\u003c/em\u003e. 2019;19(10):651\u0026ndash;667. doi:10.1038/s41568-019-0177-4.\u003c/li\u003e\n\u003cli\u003eVousden KH, Prives C. Blinded by the Light: The Growing Complexity of p53. \u003cem\u003eCell\u003c/em\u003e. 2009;137(3):413\u0026ndash;431. doi:10.1016/j.cell.2009.04.037.\u003c/li\u003e\n\u003cli\u003eKaur M, et al. Study of Ki-67 and p53 expression in cervical neoplasia. \u003cem\u003eHealthcare Bulletin\u003c/em\u003e. 2025;15(5):639\u0026ndash;645.\u003c/li\u003e\n\u003cli\u003eOgbu CC, et al. Correlation of p16, p53, Ki-67 in noninvasive cervical lesions. \u003cem\u003eJ Adv Surg Clin Res\u003c/em\u003e. 2022;12(3):25\u0026ndash;32.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-cancer","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bcan","sideBox":"Learn more about [BMC Cancer](http://bmccancer.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/bcan/default.aspx","title":"BMC Cancer","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Cervical intraepithelial neoplasia, p53, squamous cell carcinoma, immunohistochemistry, biomarker, human papillomavirus, tumor suppressor genes","lastPublishedDoi":"10.21203/rs.3.rs-7567894/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7567894/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e\u003cp\u003eThe development of cervical cancer is significantly influenced by the dysregulation of the tumor suppressor p53. While TP53 mutations are frequently present in HPV-independent malignancies, as evidenced by abnormal immunohistochemistry (IHC) patterns, E6-mediated degradation in HPV-associated illness reduces p53 function. This study examined p53 expression in a Nigerian cohort with cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC) to elucidate its function in lesion progression.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e\u003cp\u003eTwo hundred cervical specimens (CIN I\u0026ndash;III and SCC) that were archived as formalin-fixed paraffin-embedded (FFPE) were recovered. For histological confirmation, sections (3 \u0026micro;m) were stained with hematoxylin\u0026ndash;eosin (H\u0026amp;E). Then, p53 IHC was performed using the avidin\u0026ndash;biotin complex method with microwave antigen retrieval. Proportion and intensity were combined to create a semi-quantitative score (0\u0026ndash;6) for the expression. Types: low (1\u0026ndash;3), high (4\u0026ndash;6), and negative (0). In SPSS v16, associations with age and grade were examined using χ\u0026sup2;.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e\u003cp\u003eHigh levels of p53 expression were seen in 60.9% of SCC and 46.6% of CIN. CIN I (20.9%), CIN II (70.2%), CIN III (46.3%), and SCC (60.9%) are the grades. There was a significant correlation (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) between expression and lesion severity.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e\u003cp\u003ep53 expression rises with the severity of the dysplasia, indicating that it may be used as an auxiliary biomarker of the development of cervical neoplasia. Combining pattern-based p53 interpretation, p16/Ki-67 dual-stain, and HPV genotyping may improve its diagnostic and prognostic utility.\u003c/p\u003e","manuscriptTitle":"Progressive p53 Expression in the Histological Spectrum of Cervical Neoplasia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-17 05:19:30","doi":"10.21203/rs.3.rs-7567894/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2025-10-30T20:30:25+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-29T11:23:15+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"51268103196406214004008602237564976880","date":"2025-10-21T18:40:49+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-18T15:59:50+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"105342958978372893385923654544666418006","date":"2025-10-18T15:28:00+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-13T05:54:04+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"121605579655586538728848267031417867572","date":"2025-10-10T02:08:59+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"283635242217997038636327211059896158192","date":"2025-10-08T19:00:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"91082151872772486152468744300727816609","date":"2025-10-08T05:43:55+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-10-06T13:34:47+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-18T14:52:51+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-09-10T13:01:06+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-09T18:45:51+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Cancer","date":"2025-09-09T18:43:26+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-cancer","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bcan","sideBox":"Learn more about [BMC Cancer](http://bmccancer.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/bcan/default.aspx","title":"BMC Cancer","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b4504f6c-23f3-448a-8622-c816ea66a12a","owner":[],"postedDate":"October 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2025-10-17T05:19:30+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-17 05:19:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7567894","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7567894","identity":"rs-7567894","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}