不同HPV检测法及p16表达在喉鳞状细胞癌病理诊断中的应用价值

Value of different HPV detection Methods and p16 expression in pathological diagnosis of laryngeal squamous cell carcinoma

目的探讨人乳头瘤病毒(human papillomavirus,HPV)的不同检测方法及p16表达,在喉鳞状细胞癌(laryngeal squamous cell carcinoma,LSCC)病理诊断中的作用和应用价值。方法收集有完整临床病理资料的21例LSCC,按WHO(2017)头颈部肿瘤分类诊断标准,分别采用PCR-DNA反向点杂交法检测肿瘤组织中HPV-DNA,RNAscope法检测HPV 16型及18型E6、E7的mRNA,同时应用免疫组化SP法进行p16染色并分析其表达特点。结果 LSCC患者发病年龄37～62岁,中位年龄46岁;男性13例,女性8例,其中HPV 16-DNA检测5例为阳性(5/21,23. 8%),16例为阴性(16/21,76. 2%)。HPV阳性患者中1例p16强阳性(1/5,20. 0%); HPV 16 mRNA检测8例为阳性(8/21,38. 1%),其中5例同时为HPV 16-DNA阳性,3例p16阳性(3/8,37. 5%); 13例HPV 16 mRNA检测为阴性(13/21,61. 9%)。HPV阳性LSCC组织学多表现为非角化型鳞状细胞癌,肿瘤细胞呈卵圆形或梭形,细胞核深染,核仁不明显,细胞边界不清。结论 LSCC组织主要为HPV 16型,以非角化型鳞状细胞癌为主。应用RNAscope法检测HPV 16 mRNA比PCR-DNA反向点杂交法具有更高的灵敏性和特异性,p16不能作为检测LSCC组织中HPV感染状态的替代标志。

Purpose To evaluate the value of different methods in human papillomavirus (HPV) detection and p16 expression in pathological diagnosis of laryngeal squamous cell carcinoma (LSCC) tissues. Methods Twenty-one cases of LSCC with complete clinical data were collected in accordance with the new WHO classification of head and neck tumors in 2017. HPV-DNA was detected by PCR-DNA reverse dot blot hybridization,the expression of HPV 16 and 18 E6 and E7 was detected by RNAscope technology and all cases were stained with p16 by SP immunohistochemical method. The results were analyzed. Results The onset age of LSCC in this group range was 37 to 62 years,with a mean age of 46 years,including 13 males and 8 females. HPV-DNA16 was positive in 5 cases (5/21,23.8%) and 16 cases were negative (16/21,76.2%). Among the HPV-positive cases,1 case was positive for p16 (1/5,20.0%),8 cases were positive for HPV 16 mRNA (8/21,38.1%),5 of which were HPV 16-DNA positive and 3 were p16 positive (3/8,37.5%),and 13 cases of HPV 16 mRNA were negative (13/21,61.9%). The HPV-positive LSCC cases showed histologically more than non-keratinized SCC. The tumor cells were oval or fusiform,the nuclei were deeply stained,the nucleoli were not obvious,and the cell boundaries were unclear. Conclusion Type 16 is the main type of HPV in LSCC tissues. The histological type is mainly non keratinized squamous cell carcinoma. RNAscope technology is more sensitive and specific than reverse dot blot method in detecting HPV infection. p16 cannot be used as an alternative marker of HPV infection in LSCC tissues.