{"title":"福尔马林固定癌组织的HPV-16检测流程及其在口腔鳞状细胞癌中的应用","authors":"Shizuka Morodomi, Akiyuki Hirosue, Akhinur Rahman, Kyotaro Nohata, Misaki Matsuo, Omnia Reda, Samiul Alam Rajib, Haruki Saito, Hiroki Takeda, Ryoji Yoshida, Masafumi Nakamoto, Masatoshi Hirayama, Kenta Kawahara, Mitsuyoshi Takatori, Yorihisa Orita, Hideki Nakayama, Yorifumi Satou","doi":"10.1002/cam4.70544","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Background</h3>\n \n <p>Virus-related cancers are malignancies caused by specific viruses, such as human papillomavirus (HPV), hepatitis B virus, and human T-cell leukemia virus, contributing significantly to the global cancer burden through persistent infection and oncogenic transformation. The current study aimed to develop a robust HPV-16 detection method for formalin-fixed cancer specimens.</p>\n </section>\n \n <section>\n \n <h3> Materials and Methods</h3>\n \n <p>To prevent false negatives resulting from DNA fragmentation, a DNA quality check step was added. Additionally, this study used multiplex polymerase chain reaction (PCR) covering the entire HPV-16 genome to mitigate effects caused by viral sequence variation. To prove this concept, we analyzed genomic DNA extracted from oropharyngeal cancer tissues known as HPV-16-positive. Subsequently, the protocol was tested on oral squamous cell carcinoma (OSCC) samples in our cohort. Given the wide variation in HPV-16 positivity in previous studies, it remains elusive how frequently HPV-16 is positive in OSCC.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>The results showed faint bands or smears in the multiplex PCR of 7 out of 112 cases. Droplet digital PCR confirmed variable positivity levels of HPV-16, suggesting two scenarios of HPV-16 positivity in cancer tissue: cancer cells derived from infected cells or only a portion being HPV-16-positive. Finally, we comprehensively analyzed the case and identified the integration of a deleted HPV-16 genome into the intronic region of the host gene <i>TMEM94</i> on chromosome 17. To the best of our knowledge, this is the first evidence showing the integration of HPV-16 in OSCC cells and providing its complete viral sequence.</p>\n </section>\n \n <section>\n \n <h3> Conclusions</h3>\n \n <p>The established protocol should be applicable to various cancer tissues for analyzing the association with HPV-16 infection.</p>\n </section>\n </div>","PeriodicalId":139,"journal":{"name":"Cancer Medicine","volume":"14 4","pages":""},"PeriodicalIF":3.1000,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cam4.70544","citationCount":"0","resultStr":"{\"title\":\"Robust HPV-16 Detection Workflow for Formalin-Fixed Cancer Tissue and Its Application for Oral Squamous Cell Carcinoma\",\"authors\":\"Shizuka Morodomi, Akiyuki Hirosue, Akhinur Rahman, Kyotaro Nohata, Misaki Matsuo, Omnia Reda, Samiul Alam Rajib, Haruki Saito, Hiroki Takeda, Ryoji Yoshida, Masafumi Nakamoto, Masatoshi Hirayama, Kenta Kawahara, Mitsuyoshi Takatori, Yorihisa Orita, Hideki Nakayama, Yorifumi Satou\",\"doi\":\"10.1002/cam4.70544\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Background</h3>\\n \\n <p>Virus-related cancers are malignancies caused by specific viruses, such as human papillomavirus (HPV), hepatitis B virus, and human T-cell leukemia virus, contributing significantly to the global cancer burden through persistent infection and oncogenic transformation. The current study aimed to develop a robust HPV-16 detection method for formalin-fixed cancer specimens.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Materials and Methods</h3>\\n \\n <p>To prevent false negatives resulting from DNA fragmentation, a DNA quality check step was added. Additionally, this study used multiplex polymerase chain reaction (PCR) covering the entire HPV-16 genome to mitigate effects caused by viral sequence variation. To prove this concept, we analyzed genomic DNA extracted from oropharyngeal cancer tissues known as HPV-16-positive. Subsequently, the protocol was tested on oral squamous cell carcinoma (OSCC) samples in our cohort. Given the wide variation in HPV-16 positivity in previous studies, it remains elusive how frequently HPV-16 is positive in OSCC.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>The results showed faint bands or smears in the multiplex PCR of 7 out of 112 cases. Droplet digital PCR confirmed variable positivity levels of HPV-16, suggesting two scenarios of HPV-16 positivity in cancer tissue: cancer cells derived from infected cells or only a portion being HPV-16-positive. Finally, we comprehensively analyzed the case and identified the integration of a deleted HPV-16 genome into the intronic region of the host gene <i>TMEM94</i> on chromosome 17. To the best of our knowledge, this is the first evidence showing the integration of HPV-16 in OSCC cells and providing its complete viral sequence.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusions</h3>\\n \\n <p>The established protocol should be applicable to various cancer tissues for analyzing the association with HPV-16 infection.</p>\\n </section>\\n </div>\",\"PeriodicalId\":139,\"journal\":{\"name\":\"Cancer Medicine\",\"volume\":\"14 4\",\"pages\":\"\"},\"PeriodicalIF\":3.1000,\"publicationDate\":\"2025-02-20\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cam4.70544\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cancer Medicine\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cam4.70544\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"ONCOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cancer Medicine","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cam4.70544","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"ONCOLOGY","Score":null,"Total":0}
Robust HPV-16 Detection Workflow for Formalin-Fixed Cancer Tissue and Its Application for Oral Squamous Cell Carcinoma
Background
Virus-related cancers are malignancies caused by specific viruses, such as human papillomavirus (HPV), hepatitis B virus, and human T-cell leukemia virus, contributing significantly to the global cancer burden through persistent infection and oncogenic transformation. The current study aimed to develop a robust HPV-16 detection method for formalin-fixed cancer specimens.
Materials and Methods
To prevent false negatives resulting from DNA fragmentation, a DNA quality check step was added. Additionally, this study used multiplex polymerase chain reaction (PCR) covering the entire HPV-16 genome to mitigate effects caused by viral sequence variation. To prove this concept, we analyzed genomic DNA extracted from oropharyngeal cancer tissues known as HPV-16-positive. Subsequently, the protocol was tested on oral squamous cell carcinoma (OSCC) samples in our cohort. Given the wide variation in HPV-16 positivity in previous studies, it remains elusive how frequently HPV-16 is positive in OSCC.
Results
The results showed faint bands or smears in the multiplex PCR of 7 out of 112 cases. Droplet digital PCR confirmed variable positivity levels of HPV-16, suggesting two scenarios of HPV-16 positivity in cancer tissue: cancer cells derived from infected cells or only a portion being HPV-16-positive. Finally, we comprehensively analyzed the case and identified the integration of a deleted HPV-16 genome into the intronic region of the host gene TMEM94 on chromosome 17. To the best of our knowledge, this is the first evidence showing the integration of HPV-16 in OSCC cells and providing its complete viral sequence.
Conclusions
The established protocol should be applicable to various cancer tissues for analyzing the association with HPV-16 infection.
期刊介绍：
Cancer Medicine is a peer-reviewed, open access, interdisciplinary journal providing rapid publication of research from global biomedical researchers across the cancer sciences. The journal will consider submissions from all oncologic specialties, including, but not limited to, the following areas:
Clinical Cancer Research
Translational research ∙ clinical trials ∙ chemotherapy ∙ radiation therapy ∙ surgical therapy ∙ clinical observations ∙ clinical guidelines ∙ genetic consultation ∙ ethical considerations
Cancer Biology:
Molecular biology ∙ cellular biology ∙ molecular genetics ∙ genomics ∙ immunology ∙ epigenetics ∙ metabolic studies ∙ proteomics ∙ cytopathology ∙ carcinogenesis ∙ drug discovery and delivery.
Cancer Prevention:
Behavioral science ∙ psychosocial studies ∙ screening ∙ nutrition ∙ epidemiology and prevention ∙ community outreach.
Bioinformatics:
Gene expressions profiles ∙ gene regulation networks ∙ genome bioinformatics ∙ pathwayanalysis ∙ prognostic biomarkers.
Cancer Medicine publishes original research articles, systematic reviews, meta-analyses, and research methods papers, along with invited editorials and commentaries. Original research papers must report well-conducted research with conclusions supported by the data presented in the paper.
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