Lo Wan-Yu, Lai Chien-Chen, Hua Chun-Hung, Tsai Ming-Hsui, Huang Shiuan-Yi, Tsai Chang-Hai, Tsai Fuu-Jen
Department of Medical Research, China Medical University Hospital Taichung, Taiwan.
J Proteome Res. 2007 Jun;6(6):2143-51. doi: 10.1021/pr060551+. Epub 2007 Apr 24.
The purpose of this work is to differentiate between the Human papillomaviruses 18 positive (HPV18+) and negative (HPV18-) oral squamous cell carcinomas (OSCC) in oral cancer patients with cancer-associated oral habits (betel quid chewing, cigarette smoking, and alcohol drinking). Both gene and protein expression profiles of HPV18+ and HPV18- OSCC were compared: we then further explored the biological effect of HPV in oral cancer. Suppression subtraction hybridization (SSH), clinical proteomics analysis, and immunohistochemistry (IHC) staining were carried out in the HPV18+ and HPV18- OSCC groups. HPV typing detection revealed that 11 OSCC tissues from 82 patients were positive for HPV18. The SSH experiment showed that 4 cancer-associated genes were highly transcribed within 11 cDNA libraries of HPV18+ OSCC, including poly(ADP-ribose)polymerase I (PARP1), replication protein A2 (RPA2), S100A8, and S100A2. Clinical proteomics analysis indicated that there was over 10-fold overexpression of Stratifin, F-actin capping protein alpha-1 subunit (CapZ alpha-1), Apolipoprotein A-1 (ApoA-1), Heat-shock protein 27 (HSP27), Arginase-1, p16INK4A, and S100 calcium-binding protein A8 (S100A8) in HPV18+ OSCC. Interestingly, the results from SSH and protemics analysis showed that S100A8 was overexpressed in HPV18+ OSCC. Moreover, IHC staining demonstrated that S100A8 was up-regulated in HPV18+ OSCC tissues. Our results suggest that S100A8 plays an important role in oral carcinogenesis following HPV18 infection; therefore, S100A8 may be a powerful biomarker of HPV18 as well as a potential therapeutic target for HPV18+ OSCC patients. The study is the first to identify S100A8 as a biomarker in HPV-associated cancer. Furthermore, this is also the first study to discover a biomarker by combining SSH, clinical proteomics, and IHC stain analysis in oral cancer-associated research.
本研究旨在区分有癌症相关口腔习惯（嚼槟榔、吸烟和饮酒）的口腔癌患者中，人乳头瘤病毒18阳性（HPV18+）和阴性（HPV18-）的口腔鳞状细胞癌（OSCC）。比较了HPV18+和HPV18- OSCC的基因和蛋白质表达谱：然后进一步探究HPV在口腔癌中的生物学效应。对HPV18+和HPV18- OSCC组进行了抑制性消减杂交（SSH）、临床蛋白质组学分析和免疫组织化学（IHC）染色。HPV分型检测显示，82例患者中的11例OSCC组织HPV18呈阳性。SSH实验表明，在HPV18+ OSCC的11个cDNA文库中，4个癌症相关基因转录水平较高，包括聚（ADP-核糖）聚合酶I（PARP1）、复制蛋白A2（RPA2）、S100A8和S100A2。临床蛋白质组学分析表明，HPV18+ OSCC中Stratifin、F-肌动蛋白封端蛋白α-1亚基（CapZα-1）、载脂蛋白A-1（ApoA-）、热休克蛋白27（HSP27）、精氨酸酶-1、p16INK4A和S100钙结合蛋白A8（S100A8）的表达量超过10倍。有趣的是，SSH和蛋白质组学分析结果表明，S100A8在HPV18+ OSCC中过表达。此外，IHC染色显示S100A8在HPV18+ OSCC组织中上调。我们的结果表明，S100A8在HPV18感染后的口腔癌发生过程中起重要作用；因此，S100A8可能是HPV18的有力生物标志物，也是HPV18+ OSCC患者潜在的治疗靶点。该研究首次将S100A8鉴定为HPV相关癌症中的生物标志物。此外，这也是在口腔癌相关研究中首次通过结合SSH、临床蛋白质组学和IHC染色分析发现生物标志物的研究。
