Gourin Christine G, Zhi Wenbo, Adam Bao-Ling
Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University, Baltimore, Maryland 21287, USA.
Laryngoscope. 2009 Jul;119(7):1291-302. doi: 10.1002/lary.20279.
OBJECTIVES/HYPOTHESIS: Serum protein profiling by SELDI-TOF-MS distinguishes pretreatment and post-treatment samples from patients with head and neck squamous cell cancer (HNSCC) by disease status (disease-free or recurrence) with a high degree of sensitivity and specificity. We sought to identify biomarkers for recurrence with potential utility for surveillance and incorporated 2-D DIGE and MALDI-TOF-MS techniques to overcome the limitations of SELDI-TOF-MS in determining biomarker identity.
Serum samples were collected prospectively from 143 HNSCC patients and analyzed based on disease status following treatment.
Recurrent HNSCC occurred in 46 patients. MALDI-TOF-MS following immunodepletion of major plasma proteins followed by 2-D DIGE identified 181 proteins with differential expression between pretreatment and post-treatment samples collected 6 months or more following treatment. Classification by disease status revealed significant differential expression of 16 proteins, with recurrent HNSCC associated with underexpression of kininogen and serine protease inhibitors C-1 inhibitor, kininogen, angiotensinogen, serine/cysteine proteinase inhibitor clade G member 1, and overexpression of thiol-specific antioxidant proteins (TSA), apolipoprotein A1 and proapolipoprotein, and epidermal cytokeratin 2.
Serum protein profiling using 2D DIGE/MALDI-TOF-MS identifies proteins with significant differential expression in HNSCC based on disease status. Recurrent HNSCC was associated with underexpression of several protease inhibitors and kininogen, which has antiangiogenic properties, and overexpression of TSA, which is a free radical scavenger, as well as several forms of apolipoprotein A1 that may serve as a carrier molecule but may also indirectly promote tumor survival through kinase activation. This profile is consistent with a more aggressive disease variant and warrants further investigation.
目的/假设：通过表面增强激光解吸电离飞行时间质谱（SELDI-TOF-MS）进行血清蛋白谱分析，能够以高度的敏感性和特异性，根据疾病状态（无病或复发）区分头颈鳞状细胞癌（HNSCC）患者的治疗前和治疗后样本。我们试图鉴定具有复发监测潜在效用的生物标志物，并采用二维差异凝胶电泳（2-D DIGE）和基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）技术，以克服SELDI-TOF-MS在确定生物标志物身份方面的局限性。
前瞻性收集143例HNSCC患者的血清样本，并根据治疗后的疾病状态进行分析。
46例患者出现复发性HNSCC。在对主要血浆蛋白进行免疫去除后，采用MALDI-TOF-MS，随后进行2-D DIGE，鉴定出181种在治疗后6个月或更长时间收集的治疗前和治疗后样本之间存在差异表达的蛋白质。按疾病状态分类显示，16种蛋白质存在显著差异表达，复发性HNSCC与激肽原和丝氨酸蛋白酶抑制剂C1抑制剂、激肽原、血管紧张素原、丝氨酸/半胱氨酸蛋白酶抑制剂G家族成员1的低表达相关，与硫醇特异性抗氧化蛋白（TSA）、载脂蛋白A1和前载脂蛋白以及表皮细胞角蛋白2的高表达相关。
使用二维差异凝胶电泳/基质辅助激光解吸电离飞行时间质谱进行血清蛋白谱分析，可根据疾病状态鉴定出在HNSCC中存在显著差异表达的蛋白质。复发性HNSCC与几种蛋白酶抑制剂和具有抗血管生成特性的激肽原的低表达相关，与作为自由基清除剂的TSA的高表达相关，以及与几种可能作为载体分子但也可能通过激酶激活间接促进肿瘤存活的载脂蛋白A1形式相关。这种蛋白谱与一种更具侵袭性的疾病变体一致，值得进一步研究。
