广东省潮汕地区单中心头颈部鳞状细胞癌患者人乳头瘤病毒初步检测分析

目的探讨广东省潮汕地区单中心头颈部鳞状细胞癌(HNSCC)患者人乳头瘤病毒(HPV)感染及亚型状况。方法收集2014年12月至2016年12月汕头大学医学院附属肿瘤医院167例HNSCC患者的肿瘤原发灶标本。采用免疫组织化学(IHC)法检测肿瘤组织中p16蛋白的表达,以肿瘤细胞p16蛋白阳性率≥76%为判断HNSCC存在HPV的依据,分析p16蛋白与患者临床病理因素的关系。采用原位杂交法(ISH)检测肿瘤组织中是否存在HPV 16、18 DNA;应用RNAscope法检测肿瘤组织中18种常见的高危HPV亚型(HPV HR 18)RNA的表达情况,分析p16蛋白阳性细胞比例≥50%的肿瘤组织中HPV HR 18阳性情况。结果患者p16蛋白强表达率为7.2%(12/167);低龄组(<50岁)p16蛋白强表达率高于高龄组(≥50岁)17.2%(5/29)比5.1%(7/138),χ^2=5.321,P=0.021];口咽癌组p16蛋白强表达率高于非口咽癌组29.4%(5/17)比4.7%(7/150),χ^2=14.019,P<0.01];性别、烟酒嗜好及肿瘤分期分层患者间p16蛋白强表达率差异均无统计学意义(均P>0.05)。ISH检测示,全部HNSCC原发灶均未发现HPV 16、18 DNA,重复实验结果一致。RNAscope法检测示,肿瘤细胞p16蛋白阳性率≥50%的19例患者中,3例(15.8%)肿瘤组织HPV HR 18 RNA阳性。结论潮汕地区HNSCC患者的HPV阳性率较低,以口咽癌患者最高,且呈年轻化趋势。潮汕地区HNSCC主要致HPV亚型可能并非HPV 16、18,而可能是包括HPV HR 18中的其他致病亚型。

Initial analysis of human papilloma virus for patients with head and neck squamous cell carcinoma in a single center in Chaoshan area of Guangdong Province

Objective To detect the infection and subtype status of human papilloma virus(HPV)for patients with head and neck squamous cell carcinoma(HNSCC)treated in a single center in Chaoshan area of Guangdong Province.Methods The primary lesion samples from 167 HNSCC patients in Cancer Hospital of Shantou University Medical College between December 2014 and December 2016 were collected.The expression of p16 protein in tumor tissues was detected by using immunohistochemistry(IHC),and the positive rate of tumor cell p16 protein≥76%was used as a diagnostic standard to judge HPV in HNSCC;the relationship between p16 protein and the clinicopathological factors was analyzed.The status of HPV 16/18 DNA in tumor tissues was tested by using in situ hybridization(ISH).RNA scope was used to detect the RNA expression of 18 kinds of common high-risk HPV subtype(HPV HR 18),and the positive status of HPV HR 18 in tumor tissues with the positive cell proportion≥50%of p16 protein was analyzed.Results The strong expression rate of p16 protein was 7.2%(12/167).The strong expression rate of p16 protein in the younger group(<50 years old)was higher than that in the older group(≥50 years old)17.2%(5/29)vs.5.1%(7/138),χ^2=5.321,P=0.021].The strong expression rate of p16 protein in the oropharyngeal carcinoma group was higher than that in the non-oropharyngeal carcinoma group29.4%(5/17)vs.4.7%(7/150),χ^2=14.019,P<0.01].The strong expression rate of p16 protein in the gender,smoking and alcohol consumption,tumor staging and stratification among different patients was not statistically different(all P>0.05).HPV 16/18 DNA was not found in all HNSCC primary lesions by using ISH,which showed the same result after repeated examination.RNAscope method showed that 3 cases(15.8%)out of the 19 patients with p16 protein positive rate≥50%were HPV HR 18 RNA positive.Conclusions The positive rate of HPV for HNSCC patients in Chaoshan area is low,while the patients with oropharyngeal carcinoma have the highest rate and tend to be younger.The main carcinogenic viruses of HPV for HNSCC patients in Chaoshan area are other subtypes of HPV including HPV HR 18 rather than HPV 16/18.