喉鳞状细胞癌组织HPV16和p16表达的临床意义

Clinical significance of HPV16 and p16 expression in laryngeal squamous cell carcinoma

目的探讨喉鳞状细胞癌(LSCC)中人乳头瘤病毒(HPV)及细胞周期相关蛋白p16的表达及意义。方法收集北京同仁医院2005-01-01-2011-12-31LSCC病例资料332例,回顾性分析其临床病理学特点,采用聚合酶链反向点杂交(PCR-RDB)方法检测HPV16DNA,构建组织芯片并采用免疫组织化学法染色观察细胞周期相关蛋白P16的表达,比较HPV阳性组和HPV阴性组临床病理学特点的差异,分析高危型HPV16与p16的关系;采用RNAscope方法检测HPV16 E6/E7 mRNA的表达,观察在LSCC中是否具有转录活性的HPV,从而明确LSCC是否与HPV有关联。结果在332例LSCC中有45例(45/332,13.6%)检测到高危型HPV,其中40例(40/332,12.0%)检测到HPV16,HPV16阳性与淋巴结内肿瘤囊性转移有关联,χ^(2)=9.774,P=0.004;T_(1)患者HPVI6阳性率高于T_(4)患者,F=0.011;组织学分级也对患者是否表达HPV16DNA有一定的影响,P=0.049。与LSCC患者年龄、性别、是否吸烟/饮酒、TNM分期及肿瘤大小等均无统计学意义的关联,均P>0.05。在332例LSCC中36例(36/332,10.84%)p16阳性表达,其中女性阳性率高于男性,差异有统计学意义的关联,χ^(2)=4.841,P=0.04;但与年龄、是否吸烟/饮酒、TNM分期、肿瘤部位、组织学类型、组织学分级及淋巴结转移无统计学意义的关联,均P>0.05。HPVI6表达与p16表达有统计学意义的关联,χ^(2)=3.944,P=0.047。但是p16与HPV16的表达并不完全一致,其敏感性仅为20%(8/40)。332例LSCC均检测了HPV16E6/E7mRNA的表达,染色结果均为阴性。结论部分LSCC中伴有HPV16的感染,但HPV16并不是一个主要的致癌因素,且LSCC中P16表达并不是HPV16感染的替代性指标。

Objective To investigate the significance of HPV16 and cell cycle related protein p16 in LSCC.Methods All 332 LSCC cases in Beijing Tongren Hospital between 2005-01-01 and 2011-12-31 were collected and the clinicopathologic features of these patients were analyzed.HPV16 DNA were detected by polymerase chain reaction-reverse dot blot(PCR-RDB)and tissue microarray were constructed for all cases to detect the expression of cell cycle related protein p16 by immunohistochemistry.Then we compared the clinical and pathological characteristics between the HPV positive group and the HPV negative group and analyzed the relationship between HPV16 and p16.Besides,we detected the expression of HPV16 E6/E7 mRNA in LSCC by RNAscope and observe whether there was any HPV with transcriptional activity in LSCC,so as to determine whether LSCC was related with HPV.Results High risk HPV DNA were detected in 45(45/332,13.6%)of 332 LSCC and HPV16 DNA was detected in 40(40/332,12.0%).HPV16 positive was related with tumor cystic metastasis type(χ^(2)=9.774,P=0.004).The positive rate of HPV16 in T_(1)patients was higher than that in T_(4)patients(P=0.O11)and HPV16 was correlated with histological grading(P=0.049).There was no association of HPV16 with age,gender,alcohol consumption,smoking status,TNM stage or tumour size(P>0.05).Thirty-six patients(36/332,10.8%)were scored as p16 positive.Patients with p16-positivie tumours were more likely to be female(χ^(2)=4.841,P=0.04).However,there was no association of p16 with age,smoking status,alcohol consumption,TNM stage,tumour subsites,histological subtype and lymph node m etastasis(P>0.05).P16 positivity was related with HPV16 DNA expression(χ^(2)=3.944,P=0.047).The expression of p16 is not completely consistent with HPV16 and the specificity of p16 was only 20%(8/40).Totally 332 cases were detected HPV16 E6/E7 mRNA by RNAscope and none was positive.Conclusion HPV16 infection can be detected in some LSCC,but HPV16 was not a major carcinogenic factor and p16 was not surrogate marker for HPV infection in LSCC.