Snijders P J, Cromme F V, van den Brule A J, Schrijnemakers H F, Snow G B, Meijer C J, Walboomers J M
Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.
Int J Cancer. 1992 Jul 30;51(6):845-50. doi: 10.1002/ijc.2910510602.
The presence of human papillomavirus (HPV) DNA was assessed in biopsies of tonsillar carcinomas (n = 10) and cases of tonsillitis (n = 7), serving as controls, by general-primer-mediated PCR (GP-PCR). All carcinomas appeared HPV-positive, whereas all cases of tonsillitis were HPV-negative. Additional type-specific PCR for HPV 6, 11, 16, 18, 31 and 33 revealed that 4 carcinomas contained HPV 16 DNA, 4 contained HPV 33 DNA and 1 contained an HPV 16/33 double infection. False positivity was excluded by additional Southern blot analysis of type-specific PCR-positive samples (n = 4). Further characterization of GP-PCR products by sequence analysis revealed that 2 carcinomas contained still uncharacterized HPV genotypes; one of these also contained HPV 33 DNA and one was negative by type-specific PCR. Application of RNA PCR revealed expression of HPV 16 or HPV 33 E7 encoding spliced E6*1 transcripts in all tonsillar carcinomas (n = 4) examined. Additional non-radioactive RNA in situ hybridization performed on 3 biopsies revealed the presence of HPV 16 or HPV 33 E7 transcripts exclusively localized within the carcinoma cells, whereas stroma stained negative. These findings strongly support a role for certain HPV types in the pathogenesis of tonsillar carcinomas.
通过通用引物介导的聚合酶链反应（GP-PCR），在扁桃体癌活检样本（n = 10）以及作为对照的扁桃体炎病例（n = 7）中评估人乳头瘤病毒（HPV）DNA的存在情况。所有癌组织样本均显示HPV呈阳性，而所有扁桃体炎病例的样本HPV均为阴性。针对HPV 6、11、16、18、31和33的额外型特异性PCR检测显示，4例癌组织样本含有HPV 16 DNA，4例含有HPV 33 DNA，1例为HPV 16/33双重感染。通过对型特异性PCR阳性样本（n = 4）进行额外的Southern印迹分析排除了假阳性。通过序列分析对GP-PCR产物进行进一步鉴定，结果显示2例癌组织样本含有尚未鉴定的HPV基因型；其中1例还含有HPV 33 DNA，1例在型特异性PCR检测中呈阴性。RNA PCR检测显示，在所检测的所有扁桃体癌组织样本（n = 4）中均有HPV 16或HPV 33 E7编码的剪接E6*1转录本表达。对3份活检样本进行的额外非放射性RNA原位杂交显示，HPV 16或HPV 33 E7转录本仅存在于癌细胞内，而基质染色为阴性。这些发现有力地支持了某些HPV类型在扁桃体癌发病机制中所起的作用。
