Chromogenic in situ hybridization as a tool for hpv-related head and neck cancer diagnosis

Sophie Outh-Gauer, Jérémy Augustin, Marion Mandavit, Ophélie Grard, Thomas Denize, Marine Nervo, Charles Lépine, Marc Rassy, Eric Tartour, Cécile Badoual

    Résultats de recherche: Contribution à un journalArticleRevue par des pairs

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    Résumé

    Human papillomavirus (HPV) infection is a major risk factor for a subtype of oropharyngeal squamous cell carcinoma (OPSCC), which tends to be associated with a better outcome than alcohol-and tobacco-related OPSCC. Chromogenic in situ hybridization (CISH) of HPV viral RNA could allow the semiquantitative evaluation of viral transcripts of the oncogenic proteins E6 and E7 and an in situ visualization with a good spatial resolution. This technique allows the diagnosis of an active infection with the visualization of HPV transcription in the tumoral HPV-infected cells. An advantage of this technique is the avoidance of contamination from nonneoplastic HPV-infected cells adjacent to the tumor. Overall, its good diagnosis performances have it considered to be the gold standard for active HPV infection identification. Since E6 and E7 viral protein interaction with cell proteins pRb and p53 is mandatory for cell transformation, HPV RNA CISH is functionally relevant and acutely reflects active oncogenic HPV infection. This technique is clinically relevant as well since "low" or "high" HPV transcription levels helped the identification of two prognosis groups among HPV-related p16-positive head and neck cancer patients. Here we present the protocol for manual HPV RNA CISH performed on formalin-fixed paraffin-embedded (FFPE) slides with a kit obtained from the manufacturer. Instead of chromogenic revelation, RNA in situ hybridization may also be performed with fluorescent revelation (RNA FISH). It may also be combined with conventional immunostaining.

    langue originaleAnglais
    Numéro d'articlee59422
    journalJournal of Visualized Experiments
    Volume2019
    Numéro de publication148
    Les DOIs
    étatPublié - 1 juin 2019

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