Zander Kelly A, Saavedra Milene T, West James, Scapa Victor, Sanders Linda, Kingdom Todd T
Department of Otolaryngology-Head and Neck Surgery, University of Colorado Denver, Denver, Colorado, USA.
Am J Rhinol Allergy. 2009 May-Jun;23(3):268-72. doi: 10.2500/ajra.2009.23.3314.
The purpose of this study was to apply protein microarray technology to the study of sinonasal tissue and to identify differential protein expression in nasal polyps from aspirin-sensitive (AS) versus aspirin-tolerant (AT) patients with chronic rhinosinusitis (CRS) and CRS with nasal polyps (CRSwNPs).
Nasal polyp specimens were prospectively obtained from two groups of patients with CRSwNP. The test group (AS) consisted of five patients that were diagnosed with CRSwNP and intolerance to aspirin based on medical history and physical exam. The control group (AT) consisted of four AT patients with CRSwNP. Protein was extracted and labeled from harvested polyps and the Sigma Panorama Antibody Microarray-Cell Signaling Kit was used to identify differences in protein expression between the two polyp groups. Western blot analysis was used to validate the results of the protein microarray.
The protein microarray showed a greater than twofold change in expression of both beta-adaptin and heat shock protein 70 (HSP70). Western blot analysis confirmed up-regulation of beta-adaptin and HSP70 in nasal polyp tissue from AS patients.
Pooled samples of AS and AT nasal polyps evaluated by protein microarray show distinct protein expression profiles in the stress response and receptor-mediated endocytosis pathways. This study establishes the successful application of protein microarray technology to study nasal polyposis, which in turn can be validated by Western blot analysis.
本研究旨在将蛋白质芯片技术应用于鼻窦组织研究,以鉴定慢性鼻窦炎(CRS)和伴鼻息肉的慢性鼻窦炎(CRSwNP)患者中,阿司匹林敏感(AS)组与阿司匹林耐受(AT)组鼻息肉组织中的差异蛋白表达。
前瞻性地从两组CRSwNP患者中获取鼻息肉标本。试验组(AS)由5例根据病史和体格检查诊断为CRSwNP且对阿司匹林不耐受的患者组成。对照组(AT)由4例患有CRSwNP的AT患者组成。从采集的息肉中提取并标记蛋白质,使用西格玛全景抗体芯片-细胞信号转导试剂盒鉴定两组息肉组织中蛋白质表达的差异。采用蛋白质印迹分析验证蛋白质芯片的结果。
蛋白质芯片显示β-衔接蛋白和热休克蛋白70(HSP70)的表达变化均超过两倍。蛋白质印迹分析证实AS患者鼻息肉组织中β-衔接蛋白和HSP70上调。
通过蛋白质芯片评估的AS和AT鼻息肉混合样本在应激反应和受体介导的内吞途径中显示出不同的蛋白质表达谱。本研究证实了蛋白质芯片技术在鼻息肉病研究中的成功应用,该技术反过来可通过蛋白质印迹分析进行验证。