Mills Anne M, Dirks Dawn C, Poulter Melinda D, Mills Stacey E, Stoler Mark H
Department of Pathology, University of Virginia, Charlottesville, VA.
Am J Surg Pathol. 2017 May;41(5):607-615. doi: 10.1097/PAS.0000000000000800.
Dysregulated expression of oncogenic types of E6 and E7 is necessary for human papillomavirus (HPV)-driven carcinogenesis. An HPV E6/E7 mRNA in situ hybridization (ISH) assay covering 18 common high-risk types ("HR-RISH," aka HR-HPV RNA18 ISH) has not been extensively studied in the anogenital tract or validated on automated technology. We herein compare HR-RISH to DNA polymerase chain reaction (PCR), p16 immunohistochemistry, and a previously available HPV DNA ISH assay in HPV-related anogenital and head and neck (H&N) neoplasia. A total of 102 squamous intraepithelial lesions (16 CIN1, 25 CIN3, 3 AIN1, 12 AIN3, 9 VIN3)/invasive squamous cell carcinomas (17 cervical, 2 anal, 18 H&N) as well as 10 normal and 15 reactive cervix samples were collected. HR-RISH, DNA ISH, and p16 immunohistochemistry were performed on whole formalin-fixed, paraffin-embedded sections. RNA ISH for 6 low-risk HPV types (LR-RISH) was also performed. RNA and DNA ISH assays used automated systems. HR-HPV PCR was performed on morphology-directed formalin-fixed, paraffin-embedded punches. HR-RISH was ≥97% sensitive for PCR+ and p16+ neoplasia, as well as morphologically defined anogenital high grade squamous intraepithelial lesion/invasive squamous cell carcinoma. HR-RISH was also positive in 78% of anogenital low grade squamous intraepithelial lesion, including 81% of CIN1. Furthermore, a subset of PCR-negative/invalid and p16-negative lesions was positive for HR-RISH. Only 1 problematic reactive cervix sample and no normal cervix samples stained. These results demonstrate that HR-RISH is a robust method for the detection of HR-HPV-related neoplasia and provides insight into HPV pathobiology. Performance meets or exceeds that of existing assays in anogenital and H&N lesions and may play a role in resolving diagnostically challenging CIN1 versus reactive cases.
致癌型E6和E7的表达失调是人类乳头瘤病毒（HPV）驱动的致癌作用所必需的。一种涵盖18种常见高危类型的HPV E6/E7 mRNA原位杂交（ISH）检测方法（“HR-RISH”，即HR-HPV RNA18 ISH）尚未在肛门生殖道中得到广泛研究，也未在自动化技术上得到验证。我们在此将HR-RISH与DNA聚合酶链反应（PCR）、p16免疫组织化学以及先前可用的HPV DNA ISH检测方法在HPV相关的肛门生殖道和头颈部（H&N）肿瘤中进行比较。总共收集了102例鳞状上皮内病变（16例CIN1、25例CIN3、3例AIN1、12例AIN3、9例VIN3）/浸润性鳞状细胞癌（17例宫颈、2例肛门、18例H&N）以及10例正常宫颈样本和15例反应性宫颈样本。对整个福尔马林固定、石蜡包埋切片进行HR-RISH、DNA ISH和p16免疫组织化学检测。还对6种低危HPV类型进行了RNA ISH检测（LR-RISH）。RNA和DNA ISH检测使用自动化系统。对形态学导向的福尔马林固定、石蜡包埋组织块进行HR-HPV PCR检测。HR-RISH对PCR阳性和p16阳性肿瘤以及形态学定义的肛门生殖道高级别鳞状上皮内病变/浸润性鳞状细胞癌的敏感性≥97%。HR-RISH在78%的肛门生殖道低级别鳞状上皮内病变中也呈阳性，其中包括81%的CIN1。此外，一部分PCR阴性/无效和p16阴性病变的HR-RISH呈阳性。只有1例有问题的反应性宫颈样本染色，正常宫颈样本均未染色。这些结果表明，HR-RISH是检测HR-HPV相关肿瘤的一种可靠方法，并为HPV病理生物学提供了见解。其性能在肛门生殖道和H&N病变中达到或超过了现有检测方法，可能在解决诊断具有挑战性的CIN1与反应性病例方面发挥作用。
