摘要
目的研究中耳胆脂瘤中核因子-κB(nuclearfactorkappaB,NF-κB)的表达与活化,深入阐明胆脂瘤的发病机制。方法中耳手术中收集21例中耳胆脂瘤组织及8例正常外耳道皮肤组织,分别采用免疫组化及凝胶电泳迁移阻滞法(electrophoreticmobilityshiftassay,EMSA)检测2种组织NF-κB的蛋白表达分布及DNA结合活性,并进一步以胆脂瘤鳞屑刺激人角质形成细胞系HaCaT,观察细胞NF-κB的活性变化。结果胆脂瘤上皮组织中NF-κB的表达强度显著高于正常表皮组织,其阳性细胞平均积分吸光度分别为0·168±0·051、0·088±0·019(t=4·211,P<0·01),部分胆脂瘤(12/21)上皮细胞出现明显NF-κB的核转位;EMSA结果显示胆脂瘤组电泳带相对密度扫描值平均为(16·5±10·1)%,正常皮肤组则为(1·38±1·24)%,提示胆脂瘤组织中NF-κB的DNA结合活性显著高于正常皮肤组织(t=3·600,P=0·014);在胆脂瘤鳞屑刺激下,HaCaT细胞出现NF-κB的活化,其活性变化与鳞屑组织呈剂量依赖关系。结论NF-κB在胆脂瘤组织中存在异常活化,核因子-κB可能在胆脂瘤的发生和持续发展中起重要作用。
Objective To explore the expression and activation of NF-κB in middle ear cholesteatoma. Methods The protein expression of NF-κB p65 in 21 middle ear cholesteatoma tissues and 8 normal external ear canal skin obtained in middle ear surgery were examined by immunohistochemitry; NF-κB DNA binding activity in these two kinds of tissues were also detected by electrophoretic motility shift assay（EMSA）. The influence of cholesteatoma debris on the NF-κB DNA binding activity of HaCat cell were further analyzed. Results All epithelial cell of cholesteatoma revealed a relatively abundant plasm expression of NF-κB p65 protein, among which 12 cases showed nuclear positive expression. In contrast,the normal skin epithelium only revealed a sparse plasm distribution of NF-κB protein. The levels of NF-κB p65 protein expression in the epithelium of middle ear cholesteatoma tissue and normal skin were 0. 168 ± 0. 051,0. 088 ±0. 019（ t = 4. 211, P 〈 0.01 ）, respectively. The NF-κB DNA binding activities of cholesteatoma [ （ 16. 5 ± 10. 1 ） % ] were also higher than those in normal skin [ （ 1.38 ± 1.24） %, t = 3. 600, P = 0. 014 ]. The NF-κB DNA binding activity of HaCat cell increased when exposed to cholesteatoma debris in a dose dependent manner. Conclusion NF-κB might be an important factor which was involved in the occurrence and development of cholesteatoma.
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2006年第6期455-459,共5页
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
基金
湖北省科技攻关计划资助项目(2002AA301C56)
关键词
胆脂瘤
中耳
NF-ΚB
电泳
聚丙烯酰氨凝胶
免疫组织化学
Cholesteatoma, middle ear
NF-kappa B
Electrophoresis, polyacrylamide gel
Immunohistochemistry
