A novel RT-PCR method for quantification of human papillomavirus transcripts in archived tissues and its application in oropharyngeal cancer prognosis.
A novel RT-PCR method for quantification of human papillomavirus transcripts in archived tissues and its application in oropharyngeal cancer prognosis.
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一种新型的RT-PCR方法，用于定量存档组织中人乳头瘤病毒转录物及其在口咽癌预后中的应用。
DOI：
10.1002/ijc.27739
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发表时间：
2013-02-15
影响因子：
6.4
通讯作者：
Wang, Xiaowei
中科院分区：
文献类型：
--
作者：
Gao, Ge;Chernock, Rebecca D.;Gay, Hiram A.;Thorstad, Wade L.;Zhang, Tian R.;Wang, Hongwei;Ma, Xiao-Jun;Luo, Yuling;Lewis, James S., Jr.;Wang, Xiaowei
口咽鳞状细胞癌（SCC）与人乳头瘤病毒（HPV）感染密切相关，这与大多数其他头颈部癌症明显不同。然而，一种强大的定量反转录PCR （RT-qPCR）方法用于常规固定组织中HPV基因的全面表达谱分析尚未报道。为了解决这一问题，我们建立了一种新的实时RT-PCR方法，用于13种高危型HPV的E6和E7癌基因的表达谱分析。该方法在宫颈癌中得到了验证，并与口咽肿瘤中另一种HPV RNA检测方法（原位杂交）进行了比较。此外，还分析了选定的hpv相关人类基因的表达谱。然后在150个存档的口咽SCC样本中分析HPV E6和E7表达谱，并与其他变量和患者预后进行比较。我们的研究表明，RT-qPCR和RNA原位杂交在确定HPV状态方面是100%一致的。在大多数口咽鳞状细胞癌中发现了HPV转录活性（81.3%），患病率高于以往的研究。除HPV16外，还检测到其他三种HPV类型，包括33、35和18。此外，HPV和p16具有基本相同的表达特征，HPV和p16都是预测疾病结局的预后生物标志物。因此，p16 mRNA或蛋白表达特征是HPV转录活性（所有基因型组合）的敏感和特异性替代标志物。
Oropharyngeal squamous cell carcinoma (SCC) is strongly associated with human papillomavirus (HPV) infection, which is distinctively different from most other head and neck cancers. However, a robust quantitative reverse transcription PCR (RT-qPCR) method for comprehensive expression profiling of HPV genes in routinely fixed tissues has not been reported. To address this issue, we have established a new real-time RT-PCR method for the expression profiling of the E6 and E7 oncogenes from 13 high-risk HPV types. This method was validated in cervical cancer and by comparison with another HPV RNA detection method (in situ hybridization) in oropharyngeal tumors. In addition, the expression profiles of selected HPV-related human genes were also analyzed. HPV E6 and E7 expression profiles were then analyzed in 150 archived oropharyngeal SCC samples and compared with other variables and with patient outcomes. Our study showed that RT-qPCR and RNA in situ hybridization were 100% concordant in determining HPV status. HPV transcriptional activity was found in most oropharyngeal SCC (81.3%), a prevalence that is higher than in previous studies. Besides HPV16, three other HPV types were also detected, including 33, 35 and 18. Furthermore, HPV and p16 had essentially identical expression signatures, and both HPV and p16 were prognostic biomarkers for the prediction of disease outcome. Thus, p16 mRNA or protein expression signature is a sensitive and specific surrogate marker for HPV transcriptional activity (all genotypes combined).
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P16阳性口咽鳞状细胞癌：无论肿瘤HPV状态如何，具有良好预后的实体。
DOI：
10.1097/pas.0b013e3181e84652
发表时间：
2010-08
期刊：
The American journal of surgical pathology
影响因子：
--
作者：
Lewis JS Jr;Thorstad WL;Chernock RD;Haughey BH;Yip JH;Zhang Q;El-Mofty SK
通讯作者：
El-Mofty SK
DOI：
10.1086/597755
发表时间：
2009-05-01
期刊：
The Journal of infectious diseases
影响因子：
--
作者：
D'Souza G;Agrawal Y;Halpern J;Bodison S;Gillison ML
通讯作者：
Gillison ML
影响因子：
5.8
作者：
Larkin, M. A.;Blackshields, G.;Higgins, D. G.
通讯作者：
Higgins, D. G.
影响因子：
10.3
作者：
Fakhry, Carole;Westra, William H.;Gillison, Maura L.
通讯作者：
Gillison, Maura L.
DOI：
10.1056/nejmoa0912217
发表时间：
2010-07-01
期刊：
The New England journal of medicine
影响因子：
--
作者：
Ang KK;Harris J;Wheeler R;Weber R;Rosenthal DI;Nguyen-Tân PF;Westra WH;Chung CH;Jordan RC;Lu C;Kim H;Axelrod R;Silverman CC;Redmond KP;Gillison ML
通讯作者：
Gillison ML