p16^INK4a/Ki‐67 co‐expression specifically identifies transformed cells in the head and neck region

Abstract

p16INK4a immunohistochemical overexpression is an overall reliable surrogate marker of human papillomavirus (HPV)-associated head and neck squamous cell carcinomas (HNSCC High-risk human papillomavirus (HR-HPV)-induced head and neck squamous cell carcinomas (HNSCC) represent an individual tumor entity with biological and clinical characteristics that distinguish them from the classical smoking-or alcohol-related HNSCC. 1,2 The reliable identification of HPVinduced HNSCC will become important for HPV-stratified therapy protocols in clinical practice.The sole detection of HPV DNA cannot prove carcinogenic relevance of HPV in the concerned tumors. Therefore, additional biomarkers are required for the identification of a transforming HPV infection. Overexpression of the cell cycle regulator p16INK4a is a known surrogate marker of HPV E7 oncoprotein expression, and consequently of HPV-induced transformation.3 p16 INK4a immunohistochemistry (IHC) is widely applied for the detection of a transforming HPV infection at the uterine cervix.4,5 By analogy, p16 INK4a immunohistochemical overexpression also indicates HPV-induced HNSCC with overall high sensitivity and specificity, particularly within the oropharynx.6-9 However, some central aspects impede the unrestricted diagnostic use of p16 INK4aimmunochemistry for the detection of HPV-induced

Prognostic Impact of AJCC/UICC 8th Edition New Staging Rules in Oropharyngeal Squamous Cell Carcinoma

“…For detection, DNA was extracted from variable numbers of FFPE tissue sections depending on the tissue size (10 µm sections, approximately corresponding to 10 mm × 10 mm tumor tissue) using the DNeasy Blood & Tissue Kit by Qiagen, Hilden, Germany, according to the manufacturer’s instructions. Extracted DNA was analyzed for mucosal high-risk HPV DNA and HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 73, and 82) by PCR optimized for DNA extracted from FFPE tissue, followed by bead-based hybridization (Luminex Technology, Multimetrix, Progen, Heidelberg), as described previously (20, 21). HPV-DNA contamination protection steps (e.g., new microtome blades for every block, separated pre- and post-PCR areas) were applied and appropriate controls (tissue-free paraffin blocks, water controls) were processed to monitor potential HR–HPV DNA contamination.…”

CD56-positive lymphocyte infiltration in relation to human papillomavirus association and prognostic significance in oropharyngeal squamous cell carcinoma

“…Extracted DNA was analyzed for mucosal high risk HPV-DNA and HPV genotypes (16,18,31,33,35,39,45, 51, 52, 56, 58, 59, 68, 73 and 82) by PCR optimized for DNA extracted from FFPE tissue, followed by bead-based hybridization (Luminex technology, Multimetrix, Progen, Heidelberg), as described previously. 17,18 HPV-DNA contamination protection steps (e.g., new microtome blades for every block, separated pre and post PCR areas) were applied and appropriate controls (tissue-free paraffin blocks, water-controls) were processed to monitor potential high risk HPV-DNA contamination. Amplification of b-globin was used to ensure DNA integrity.…”

Matrix metalloproteinase (MMP) and immunosuppressive biomarker profiles of seven head and neck squamous cell carcinoma (HNSCC) cell lines

“…Without p16 Ink4a there is inappropriate cell division and cell proliferation, which is why it is not surprising to find our immortalized cancer cell lines lacking p16 Ink4a . However, p16 Ink4a overexpression is sometimes used as a surrogate marker for high-risk-HPV-associated HNSCC (24,41,42). This is because the E7 HPV oncogene protein will induce p16 Ink4a expression.…”