Department of Otolaryngology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University Hospital, Taiwan.
Laryngoscope. 2012 Dec;122(12):2818-23. doi: 10.1002/lary.23547.
OBJECTIVES/HYPOTHESIS: The mechanisms of cholesteatoma proliferation and growth remain unclear. The objective of this study is to discover the potential mechanisms of the proliferation and growth of cholesteatoma by direct experimental assessments on cholesteatoma tissues from patients.
Retrospective study by the comparisons between cholesteatoma tissues and retroauricular skin tissues from the patients.
Two-dimensional gel electrophoresis, LC-MS/MS analysis and immunohistochemistry were performed to investigate specific protein expression in cholesteatoma tissues compared with retroauricular skin tissues collected from the patients. Western blotting analysis was conducted to verify the regulation of specific proteins found by 2-DE, and to determine the changes of associated potential modulators in cholesteatoma proliferation and growth.
Twelve serpin B3 isoforms were found by 2-DE and identified by LC-MS/MS analysis, which is coherent with the results exhibited by immunohistochemistry and western blot. Up-regulation of STAT3 and down-regulations of cathepsin K and cathepsin L were represented using western blot.
The data in this study suggested serpin B3, STAT3, cathepsin K, and cathepsin L are associated with the proliferation and growth of cholesteatoma, and these proteins may be influential factors in cholesteatoma growth.
目的/假设:胆脂瘤增殖和生长的机制仍不清楚。本研究的目的是通过对患者胆脂瘤组织的直接实验评估,发现胆脂瘤增殖和生长的潜在机制。
通过对患者的胆脂瘤组织和耳后皮肤组织进行比较的回顾性研究。
采用二维凝胶电泳、LC-MS/MS 分析和免疫组织化学方法,研究胆脂瘤组织与从患者收集的耳后皮肤组织中特定蛋白的表达差异。进行 Western 印迹分析以验证 2-DE 发现的特定蛋白的调节,并确定与胆脂瘤增殖和生长相关的潜在调节剂的变化。
通过 2-DE 发现了 12 种丝氨酸蛋白酶抑制剂 B3 同种型,并通过 LC-MS/MS 分析进行了鉴定,这与免疫组织化学和 Western 印迹的结果一致。Western 印迹显示 STAT3 上调,组织蛋白酶 K 和组织蛋白酶 L 下调。
本研究的数据表明丝氨酸蛋白酶抑制剂 B3、STAT3、组织蛋白酶 K 和组织蛋白酶 L 与胆脂瘤的增殖和生长有关,这些蛋白质可能是胆脂瘤生长的影响因素。