Department of Pathology and Immunology, Washington University, 660 S. Euclid Ave., St. Louis, MO 63110, USA.
Am J Surg Pathol. 2011 Sep;35(9):1343-50. doi: 10.1097/PAS.0b013e318220e59d.
Human papillomavirus (HPV) is established as causative in oropharyngeal squamous cell carcinomas (OSCCs), being detected in 50% to 80% of tumors by DNA in situ hybridization (ISH) and/or polymerase chain reaction. However, these tests do not assess viral transcription. Many consider E6/E7 messenger ribonucleic acid (mRNA) the best indicator of HPV status, but it has not been detected in situ in OSCC. We constructed tissue microarrays (TMAs) from a cohort of OSCC for which p16 immunohistochemistry and HPV DNA ISH were previously performed on whole sections. We utilized a novel, chromogenic RNA ISH assay called RNAscope to detect E6/E7 mRNA of HPV-16 and other high-risk types on these TMAs. RNA ISH results were obtained for 196 of 211 TMA cases, of which 153 (78.1%) were positive. p16 immunohistochemistry and HPV DNA ISH were positive in 79.0% and 62.4% of cases, respectively. Concordance between RNA and p16, DNA and p16, and RNA and DNA were 96.4%, 78.7%, and 83.5%, respectively. Only 7 cases (3.6%) were discrepant between RNA ISH and p16. In univariate analysis, all 3 tests correlated with better overall survival (OS), disease-specific survival (DSS), and disease-free survival (DFS) (all P<0.001). In multivariate analysis, OS correlated significantly with RNA (hazard ratio=0.39, P=0.001), DNA (0.53, P=0.03), and p16 (0.30, P<0.001), but DSS and DFS correlated significantly only with p16 (DSS: 0.36, P=0.006; DFS: 0.42, P=0.016). RNA ISH is more sensitive than DNA ISH in detecting HPV in OSCC, and it correlates strongly with p16. Although both tests were comparable, p16 more strongly stratified patient outcomes.
人乳头瘤病毒（HPV）已被确定为口咽鳞状细胞癌（OSCC）的致病因素，通过原位杂交（ISH）和/或聚合酶链反应，在 50%至 80%的肿瘤中检测到 HPV。然而，这些检测并不能评估病毒转录。许多人认为 E6/E7 信使核糖核酸（mRNA）是 HPV 状态的最佳指标，但在 OSCC 中尚未原位检测到。我们构建了一组 OSCC 的组织微阵列（TMA），此前对这些 OSCC 的全切片进行了 p16 免疫组化和 HPV DNA ISH 检测。我们利用一种新的、显色 RNA ISH 检测方法，称为 RNAscope，在这些 TMA 上检测 HPV-16 和其他高危型别的 E6/E7 mRNA。对 211 例 TMA 中的 196 例获得了 RNA ISH 结果，其中 153 例（78.1%）为阳性。p16 免疫组化和 HPV DNA ISH 的阳性率分别为 79.0%和 62.4%。RNA 与 p16、DNA 与 p16、以及 RNA 与 DNA 的一致性分别为 96.4%、78.7%和 83.5%。只有 7 例（3.6%）在 RNA ISH 与 p16 之间存在差异。在单变量分析中，所有 3 项检测均与更好的总生存期（OS）、疾病特异性生存期（DSS）和无病生存期（DFS）相关（均 P<0.001）。在多变量分析中，OS 与 RNA（风险比=0.39，P=0.001）、DNA（0.53，P=0.03）和 p16（0.30，P<0.001）显著相关，但 DSS 和 DFS 仅与 p16 显著相关（DSS：0.36，P=0.006；DFS：0.42，P=0.016）。RNA ISH 比 DNA ISH 更敏感地检测 OSCC 中的 HPV，并且与 p16 强烈相关。虽然两种检测方法的效果相当，但 p16 更能分层患者的预后。
