口咽鳞癌组织中人乳头状瘤病毒DNA状态与p16蛋白表达的相互关系及临床意义

Relationship between HPV-DNA status and p16 protein expression in oropharyngeal squamous cell carcinoma and their clinical significance

目的探讨联合检测口咽鳞癌组织标本中人乳头状瘤病毒（HPV）的感染情况与p16蛋白表达的临床意义。方法选择66例口咽鳞癌活榆或手术标本，采用“三明治”技术对组织标本进行切片，以SPF10-DNA LiPA分型方法检测HPV—DNA状态，采用免疫组织化学法检测p16蛋白的表达情况，分析不同HPV—DNA和p16蛋白表达患者的预后。结果66例口咽鳞癌患者中，HPV—DNA阳性11例，阳性率为16．7％（11／66）。11例HPV．DNA阳性患者中，p16蛋白阳性9例，阴性2例。55例HPV—DNA阴性患者中，p16蛋白阳性12例，阴性43例。在口咽鳞癌中，HPV—DNA的表达与p16蛋白的表达具有显著的相关性（P〈0．001）。HPV—DNA和p16蛋白均为阳性组（A组）9例，HPV—DNA阳性／p16蛋白阴性或HPV—DNA阴性／p16蛋白阳性组（B组）14例，HPV—DNA和p16蛋白均为阴性组（C组）43例。A组、B组和C组患者的3年总生存率分别为100％、77．8％和42．0％，差异有统计学意义（P=0．001）；3年疾病特异性生存率分别为100％、77．8％和46．4％，差异有统计学意义（P=0．004）。结论在口咽鳞癌中，HPV—DNA的表达与p16蛋白的表达具有显著的相关性，p16蛋白是较为可靠的预测HPV状态的替代标志。联合检测HPV—DNA和p16蛋白的状态，有助于更加准确地对口咽鳞癌进行分层，并判断预后。

Objective To investigate the relationship between HPV-DNA status and p16 protein expression in oropharyngeal squamous cell carcinoma （OSCC） and their clinical significance. Methods Sixty-six patients with oropharyngeal squamous cell carcinomas treated in the Cancer Hospital of Chinese Academy of Medical Sciences from Jan. 1999 to Dec. 2009 were included in this study. Their formalin-fixed and paraffin-embedded tumor tissue blocks met the eligibility criteria and were used in this study. A ＂sandwich＂ technique was used to prepare paraffin sections for HPV-DNA analysis. HPV-DNA was detected using the SPF10 LiPA25 version 1 assay. The expression of p16 protein was detected by immunohistochemistry. The survival rates of patients with different HPV-DNA and p16 protein status were analyzed. Results HPV-DNA was detected in 11 （16.7%） of all specimens. Expression of p16 protein was detected in 9 of the 11 patients with HPV-positive tumors, and in 12 patients of 55 HPV-negative tumors. The expression of p16 protein was highly correlated with the presence of HPV-DNA （P 〈 0.001 ）. The tumors were classified into three groups based on the p16 protein expression and HPV-DNA status： group A （9 patients） ： HPV（ ＋ ） and p16 protein（ ＋ ） ; group B （ 14 patients） ： HPV-DNA（ ＋ ）/p16 protein（ - ） or HPV-DNA（ - ）/p16 protein（ ＋ ） ; and group C （43 patients） ： HPV-DNA（ - ）/p16 protein（ - ）. The 3-year OS rates of these 3 groups were 100% , 77.8% and 42.0% （P = 0. 001 ） , and their DSS rates were 100% , 77.8% and 46.4% , respectively（P = 0. 004）. Conclusions In oropharyngeal squamous cell carcinomas, p16 protein expression is highly correlated with the presence of HPV-DNA, and might be a surrogate marker for HPV-positive OSCC. Combination of p16 protein and HPV-DNA status detection may help to more accurately stratify oropharyngeal carcinomas and predict their prognosis.