Klussmann Jens P, Weissenborn Soenke J, Wieland Ulrike, Dries Volker, Eckel Hans E, Pfister Herbert J, Fuchs Pawel G
Department of Oto-Rhino-Laryngology, Head and Neck Surgery, University of Cologne, 50924 Cologne, Germany.
Med Microbiol Immunol. 2003 Aug;192(3):129-32. doi: 10.1007/s00430-002-0126-1. Epub 2002 Sep 14.
Human papillomavirus (HPV) infections are thought to be one of the causal factors in the development of head and neck squamous cell carcinomas (HNSCC), particularly in tumors arising from the Waldeyer's tonsillar ring. We screened 98 carefully stratified HNSCC and different control tissues for the presence of HPV DNA by nested polymerase chain reaction (PCR) specific for genital- and Epidermodysplasia verruciformis (EV)-associated HPVs and by HPV16-specific single step PCR. Typing was performed by direct sequencing and/or sequencing of cloned amplimers. On average HNSCC showed rather low HPV DNA prevalences; 18% of the oral cavity cancers, 8% of nasopharyngeal cancers, 25% of hypopharyngeal cancers and 7% of laryngeal cancers were HPV DNA positive. In contrast, HPV sequences could be detected in 45% of the oropharyngeal cancers, particularly tonsillar carcinomas (58%). Tonsillar carcinomas were significantly more likely to be HPV positive than tumors from any other site ( P<0.001). All tonsillar cancers contained oncogenic HPV types, predominantly HPV16 (13 of 14; 93%). Unaffected tonsils were available from two of these patients, but both tested negative for HPV DNA. Furthermore, no HPV DNA could be found in tonsillar biopsy specimens from control groups. Localization and load of HPV DNA was determined in HPV16-positive tonsillar carcinomas, their metastases and in unaffected mucosa using laser-assisted microdissection and subsequent real time fluorescence PCR. We demonstrated that the HPV genome is located in the cancer cells, whereas the infection of normal mucosa is a rare event. Quantification of HPV16 DNA in samples of seven patients yielded viral loads from 6 to 153 HPV DNA copies per beta-globin gene copy and the load values in both locations were roughly comparable. These loads are comparable with data shown for other HPV-associated lesions. Statistical evaluation of data related to clinicopathological parameters showed a significant correlation of the HPV positivity of tonsillar carcinomas with tumor grading ( P=0.008) and alcohol consumption ( P=0.029). Taken together our findings show a preferential association of HPV DNA with tonsillar carcinomas. Furthermore our results argue for HPV-positive tonsillar carcinomas representing a separate tumor entity, which is less dependent on conventional HNSCC risk factors.
人乳头瘤病毒（HPV）感染被认为是头颈部鳞状细胞癌（HNSCC）发生的病因之一，尤其是在起源于瓦尔代尔扁桃体环的肿瘤中。我们通过针对生殖器和疣状表皮发育不良（EV）相关HPV的巢式聚合酶链反应（PCR）以及HPV16特异性单步PCR，对98例经过仔细分层的HNSCC组织和不同的对照组织进行了HPV DNA检测。通过直接测序和/或对克隆扩增子进行测序来进行分型。平均而言，HNSCC的HPV DNA阳性率相当低；口腔癌的阳性率为18%，鼻咽癌为8%，下咽癌为25%，喉癌为7%。相比之下，在45%的口咽癌中可检测到HPV序列，尤其是扁桃体癌（58%）。扁桃体癌HPV阳性的可能性显著高于其他任何部位的肿瘤（P<0.001）。所有扁桃体癌均含有致癌性HPV类型，主要为HPV16（14例中的13例；93%）。其中两名患者有未受影响的扁桃体，但HPV DNA检测均为阴性。此外，在对照组的扁桃体活检标本中未发现HPV DNA。使用激光辅助显微切割及随后的实时荧光PCR，在HPV16阳性的扁桃体癌及其转移灶以及未受影响的黏膜中确定HPV DNA的定位和载量。我们证明HPV基因组位于癌细胞中，而正常黏膜感染则较为罕见。对7例患者样本中的HPV16 DNA进行定量分析，每β-珠蛋白基因拷贝的病毒载量为6至153个HPV DNA拷贝，两个部位的载量值大致相当。这些载量与其他HPV相关病变的数据相当。对与临床病理参数相关的数据进行统计评估显示，扁桃体癌的HPV阳性与肿瘤分级（P=0.008）和饮酒（P=0.029）显著相关。综上所述，我们的研究结果表明HPV DNA与扁桃体癌存在优先关联。此外，我们的结果支持HPV阳性的扁桃体癌代表一种独立的肿瘤实体，其对传统HNSCC危险因素的依赖性较小。
