热休克蛋白70在中耳胆脂瘤中的异常表达

目的研究热休克蛋白(HSP70)在中耳胆脂瘤组织中的异常表达,探讨HSP70与胆脂瘤的关系及其作用.方法采用免疫组化SABC染色法、计算机图像定量分析法,对25例中耳胆脂瘤和10例正常外耳道皮肤中HSP70的表达情况进行观察.结果在25例胆脂瘤上皮组织的全层及上皮下结缔组织中炎性细胞和成纤维细胞的胞浆中均有HSP70强表达,部分核中也有强表达.10例正常外耳道皮肤中HSP70为弱表达.计算机图象分析仪定量分析,结果显示胆脂瘤上皮中HSP70阳性细胞的平均光密度为0.3309±0.0070,正常外耳道上皮为0.1798±0.0020,胆脂瘤上皮中HSP70的含量显著高于外耳道皮肤(P＜0.001).结论HSP70可能参与了胆脂瘤的形成和发展.     

Ras蛋白在中耳胆脂瘤上皮中的表达

探讨Ｒａｓ蛋白在中耳继发性胆脂瘤上皮中的表达,分析其与胆脂瘤上皮增生调节中的可能作用。方法应用免疫组化ＳＰ染色法和计算机图像分析法,检测２２例胆脂瘤上皮和１０例正常外耳道皮肤中Ｒａｓ蛋白的表达情况。     

Caspase-9和Livin在中耳胆脂瘤中的表达及意义

目的研究凋亡相关蛋白Caspase-9和Livin在中耳胆脂瘤上皮组织中的表达,揭示二者在中耳胆脂瘤发生发展中的作用。方法采用免疫组化法检测凋亡促进蛋白Caspase-9和凋亡抵制蛋白Livin在23例中耳胆脂瘤及10例正常外耳道皮肤组织中的表达,对染色结果进行量化处理和统计学分析。结果 Caspase-9和Livin蛋白在中耳胆脂瘤上皮的表达均高于在正常外耳道皮肤中的表达(P<0.01);二者在胆脂瘤中的表达呈正相关,相关系数r=0.848(P<0.05)。结论凋亡促进蛋白Caspase-9和凋亡抑制蛋白Livin在中耳胆脂瘤中的表达同步升高,提示就凋亡过程而言,二者达到了新的平衡。     

ki-67、细胞周期蛋白依赖性激酶4在胆脂瘤型中耳乳突炎中的表达及意义

目的探讨ki-67、细胞周期蛋白依赖性激酶4(CDK4)在中耳继发胆脂瘤上皮中的表达,研究它们在胆脂瘤型中耳乳突炎中的作用.方法利用免疫组化的方法,检察32例胆脂瘤上皮和11例正常外耳道皮肤中的ki-67、CDK4蛋白的表达情况.结果ki-67在胆脂瘤上皮和外耳道皮肤中均为阳性表达,阳性细胞率分别为33.2%±13.2%、14.5%±4.9%,两组之间的差异具有显著性(P＜0.05);CDK4在胆脂瘤上皮和外耳道皮肤中阳性细胞率分别为35.6%±14.6%、13.6%±5.1%,两组之间的差异具有显著性(P=0.001).结论ki-67和CDK4在胆脂瘤上皮表达增高.     

金属硫蛋白2及白细胞介素1α和6在中耳胆脂瘤组织中的表达及意义

目的 观察金属硫蛋白2( metallothionein 2,MT-2)及白细胞介素1α(IL-1α)、白细胞介素6(IL-6)在中耳胆脂瘤组织中的表达及相关性,探讨其在中耳胆脂瘤发生发展过程中的作用.方法 采用免疫组化和反转录聚合酶链反应(RT-PCR)技术检测25例中耳胆脂瘤组织和7例正常外耳道皮肤组织中MT-2及IL-1α、IL-6蛋白和MT-2 mRNA的表达情况并分析其相关性.进一步以胆脂瘤鳞屑刺激人角质细胞系HaCaT,观察细胞MT-2 mRNA和蛋白质的表达变化.结果 MT-2、IL-1α及IL-6在中耳胆脂瘤中的表达明显高于正常外耳道皮肤,差异具有统计学意义(P值均＜0.05);MT-2mRNA在中耳胆脂瘤和正常外耳道皮肤中的表达差异具有统计学意义(t=15.38,P＜0.05).中耳胆脂瘤组织中MT-2与IL-1α的表达呈正相关关系(r=0.856,P＜0.05),与IL-6的表达亦呈正相关关系(r=0.714,P＜0.05).在胆脂瘤鳞屑刺激下,HaCaT细胞MT-2的mRNA和蛋白质表达均有明显增强,其表达变化与鳞屑组织呈浓度依赖关系.结论 MT-2和IL-1α、IL-6的异常表达可能在中耳胆脂瘤的发生、发展过程中起一定的作用.胆脂瘤鳞屑组织可能通过促进MT-2的过量表达而参与了胆脂瘤上皮的过度增殖.     

端粒酶催化亚单位和c-Myc蛋白在中耳胆脂瘤上皮中的表达及意义

目的:观察端粒酶催化亚单位(TERT)和癌基因cMyc的表达产物(cMyc蛋白)在中耳胆脂瘤上皮中的表达情况,分析其在胆脂瘤上皮增殖演变过程中的作用。方法:应用SP法和计算机图像分析系统,检测TERT、cMyc蛋白在32例胆脂瘤组织和14例胆脂瘤患者外耳道皮肤中的表达情况,并进行比较。结果:TERT和cMyc蛋白在所有胆脂瘤上皮各层细胞均存在较强表达,外耳道皮肤表达较弱。各组间胆脂瘤上皮各指标平均阳性细胞率和平均吸收度(平均灰度的倒数),均高于外耳道皮肤,差异有统计学意义(P<0.05);皮下结缔组织伴有大量的炎症细胞,如:淋巴细胞、浆细胞、中性粒细胞和巨噬细胞;胆脂瘤上皮中TERT和cMyc蛋白之间存在正相关关系(P<0.01)。结论:TERT和cMyc蛋白在胆脂瘤上皮的表达从不同方面反映了胆脂瘤上皮的过度增生和角化,二者在胆脂瘤的发病过程中有协同作用;上皮下炎症反应可能是角化细胞过度增殖的一个原因。     

Caspase-8在中耳胆脂瘤上皮中的表达及意义

目的 研究caspase-8(胱天蛋白酶-8)在中耳胆脂瘤上皮的表达,探讨其在中耳胆脂瘤发病机制中的作用.方法 分别采用免疫组化法和Western Blot(蛋白质印迹法)技术检测caspase-8在36例中耳胆脂瘤组织及20例正常外耳道皮肤组织中的表达情况.结果 caspase-8在36例胆脂瘤上皮各层细胞中均有强表达,而正常外耳道皮肤表达较弱,caspase-8在胆脂瘤上皮中的蛋白表达水平较正常外耳道皮肤显著增高,差异具有统计学意义(P<0.01).结论 胆脂瘤上皮中caspase-8表达较外耳道皮肤显著增高,其可能参与了中耳胆脂瘤上皮细胞的过度凋亡及增殖的调控.     

凋亡抑制蛋白Livin及Survivin在中耳胆脂瘤上皮的表达及意义

目的探讨凋亡抑制蛋白Livin、Survivin在中耳胆脂瘤上皮的表达及与细胞凋亡的关系。方法应用免疫组织化学二步法检测25例中耳胆脂瘤标本(实验组)及15例外耳道正常皮肤组织(对照组)中Livin及Sur-vivin的表达;采用末端转移酶介导的原位缺口末端标记染色(TUNEL)技术检测Livin及Survivin在两种组织中的表达及细胞的凋亡情况。结果与正常外耳道皮肤组织相比,Livin及Survivin蛋白在胆脂瘤上皮的表达均明显下调(P<0.01),细胞凋亡显著高于正常上皮组织(P<0.01);Livin及Survivin的表达与凋亡呈负相关。结论 Livin及Survivin在中耳胆脂瘤中的表达明显下调,可能与胆脂瘤上皮细胞凋亡有关。     

凋亡抑制蛋白Survivin及Bcl-2在中耳胆脂瘤上皮中的表达及意义

目的研究凋亡抑制蛋白Survivin及Bcl-2在中耳胆脂瘤上皮中的表达及相互关系,探讨其表达对胆脂瘤增殖能力的影响。方法运用免疫组织化学SP法检测21例胆脂瘤标本及11例外耳道骨部正常皮肤组织中Survivin、Bcl-2的表达。结果Survivin、Bcl-2在胆脂瘤上皮的表达与正常外耳道皮肤上皮比较,不仅范围广,基底上层也有表达,而且表达水平显著增高,两者比较有统计学意义(P<0.01)。胆脂瘤上皮中Survivin表达指数与Bcl-2表达指数呈正相关(r=0.553,P<0.01)。结论凋亡抑制蛋白Survivin,Bcl-2在中耳胆脂瘤的异常表达可能在胆脂瘤的发生、发展过程中起重要作用,它们可能参与胆脂瘤上皮的凋亡调控过程,因此适当控制Survivin,Bcl-2的表达可能会更有效地控制中耳胆脂瘤的发展。     

Ras 蛋白在中耳胆脂瘤上皮中的表达

目的探讨Ｒａｓ蛋白在中耳继发性胆脂瘤上皮中的表达,分析其与胆脂瘤上皮增生调节中的可能作用。方法应用免疫组化ＳＰ染色法和计算机图像分析法,检测２２例胆脂瘤上皮和１０例正常外耳道皮肤中Ｒａｓ蛋白的表达情况。结果胆脂瘤上皮各层细胞均存在Ｒａｓ蛋白的较强表达,其中１５／２２为胞膜着色,５／２２胞膜胞浆均着色,２例仅胞浆着色,正常外耳道表皮中Ｒａｓ蛋白主要表达于胞膜,以基底层显色为主;两种组织Ｒａｓ蛋白阳性表达的平均积分吸光度分别为０．８７０和０．４６３,差异呈高度显著性。结论中耳继发性胆脂瘤上皮中存在Ｒａｓ蛋白的高表达,说明胆脂瘤上皮具有高度增生性。     

表皮生长因子受体和Ki67及p16在成人中耳胆脂瘤中的表达

目的:研究表皮生长因子受体(EGFR)、Ki67、p16在成人中耳继发性胆脂瘤上皮中的表达情况,分析它们之间的相互关系,探讨其表达对胆脂瘤上皮侵袭能力的影响。方法:应用免疫组织化学SP染色方法检测EGFR、Ki67和p16在30例成人中耳胆脂瘤上皮、21例成人胆脂瘤患者外耳道正常皮肤、17例正常人外耳道中的表达情况,应用计算机图像分析系统对其阳性表达进行定量分析。结果:EGFR、Ki67、p16在成人中耳继发性胆脂瘤上皮中阳性表达率分别为70.0%,60.0%,46.7%,与外耳道正常皮肤相比表达均差异有统计学意义。成人中耳胆脂瘤中EGFR、Ki67与p16之间表达均无相关性(均P>0.05)。胆脂瘤侵袭能力与EGFR、Ki67表达有显著相关性(均P<0.01)。EGFR、Ki67表达灰度值越低,表达密度越高,胆脂瘤侵袭能力越强。p16在成人中耳胆脂瘤中的表达与侵袭能力之间无相关性(P>0.05)。EGFR、Ki67、p16在成人中耳胆脂瘤中阳性细胞主要分布于上皮全层,以基底层和棘层为著,呈高度表达;而在对照组中阳性细胞仅在基底层表达,呈弱表达。结论:EGFR、Ki67、p16在成人中耳胆脂瘤中呈高表达,EGFR、Ki67的表达与成人中耳胆脂瘤的侵袭能力有高度相关性,提示成人中耳胆脂瘤具有高度增殖能力,其中细胞因子EGFR、Ki67、p16起到重要的作用。     

Expression patterns of p27Kip1 and Ki-67 in cholesteatoma epithelium

OBJECTIVES: The cell cycle must be involved in cell proliferation of the epithelium of middle ear cholesteatoma Cyclins and cyclin-dependent kinase (CDK) complexes have important regulatory roles during cell cycle progression. Cyclin-CDK complexes are in turn regulated by the cyclin-dependent kinase inhibitors (CDKIs), which generally inhibit cell cycle progression. One of the important CDKI members is p27(Kip1). The goal of this study is to evaluate the expression of p27(Kip1) and Ki-67, a proliferation marker, in cholesteatoma and in the skin of the external ear canal. METHODS: The expressions of p27(Kip1) and Ki-67 in cholesteatoma epithelium (n = 20) and ear canal epithelium (n = 7) were investigated by an immunohistochemical technique. RESULTS: In cholesteatoma epithelium specimens, the expression of p27(Kip1) was observed from the parabasal layer to the granular layer, but not in the basal layer. Ki-67 was expressed dominantly in the basal and parabasal cell layers. Their expressions tend to be increased compared with their expressions in the normal ear canal skin. The expression pattern of the proliferation marker Ki-67 in the epithelial layers of two groups was inversely related to the expression of p27(Kip1). CONCLUSIONS: In cholesteatoma, the expressions of CDKI and Ki-67 were both increased in this study. The ability to inhibit proliferative activity was also increased in the cholesteatoma epithelium. The expression pattern of the proliferation marker Ki-67 in the epithelial layers was inversely related to the expression of p27(Kip1). Not only is the proliferation activity increased, but also the ability to inhibit hyperproliferation is increased in the cholesteatoma epidermis. Despite increased proliferative activity in the cholesteatoma epidermis, epithelial cells still retain the capability to prevent cell cycle arrest by means of p27(Kip1).     

转化生长因子β1和即刻早期基因及p27在中耳胆脂瘤上皮中的表达

OBJECTIVES: To analyze the expression of transforming growth factor-beta1 (TGF-beta1), cyclin dependent kinase inhibitor (p27) and c-fos in human middle ear cholesteatomas and to investigate the correlation between their expression and the ability of erosion of cholesteatoma. METHODS: Immunohistochemical staining (SP method) of 31 cholesteatomas and 11 external ear canal skin samples from patients and 10 external ear canal skin samples from healthful men which were taken intraoperatively, was performed for c-fos, TGF-beta1 and p27 positivity. The signals representing the expression of c-fos, TGF-beta1 and p27 were observed under microscope and scanned into a computer by an image scanner. The gray-scale of positive signals were quantitated by image processing computer. RESULTS: The percentage of positive expression of TGF-beta1 and c-fos in cholesteatoma were 87.1% and 83.9%, respectively. Their expression tended to be increased greatly compared with which in the skins of the control groups. Positive p27 signals were not observed in cholesteatomas and external ear skin tissues. It showed statistically significant correlation between expression of c-fos and the ability of erosion of cholesteatoma. There was correlation between the express ion of TGF-beta1 and the ability of erosion of cholesteatoma too. But there was no correlation between the expression of c-fos and TGF-beta1. CONCLUSION: The expression of c-fos in cholesteatoma was significally higher compared with which in the skin of external auditory of cholesteatoma patients and healthful men, which indicate that c-fos plays an important role in the hyperproliferative of cholesteatoma. The expression of TGF-beta1 was significant higher in cholesteatoma, which indicate that cytokines such as TGF-beta1 play a great role in the etiology of cholesteatoma.     

继发性胆脂瘤中CCL27的定位及检测

目的 明确趋化因子CCL27在继发性中耳胆脂瘤中的病理学表达,探讨可能的临床意义。 方法 应用免疫组织化学SP法检测CCL27在继发性中耳胆脂瘤(n=30)及正常耳后皮肤组织(n=30)的表达情况,并分析该趋化因子与胆脂瘤病理学之间的联系。 结果 继发性胆脂瘤、耳后皮肤组织中CCL27的阳性率分别为6.7%和33.3%,差异有统计学意义(P=0.021)。 结论 继发性胆脂瘤组织中CCL27的低表达可能与胆脂瘤的增殖过于活跃有关,CCL27可能在继发性中耳胆脂瘤发病过程中扮演重要角色。     

Increased expression of p63 and survivin in cholesteatomas

Conclusion. This study showed increased expression of p63 and survivin in cholesteatoma. Our finding indicates a putative role of p63 and survivin in the development of certain cholesteatomas. Objectives. Keratinocytes in cholesteatoma demonstrate uncoordinated hyperproliferation, migration, and invasion properties. p63 is a p53 homologue and a marker expressed in replicating keratinocytes. Survivin is an inhibitor of apoptosis protein that is abundantly expressed in most solid and hematologic malignancies. The purpose of this study was to investigate the differential expression of p63 and survivin in human middle ear cholesteatoma epithelium. Materials and methods. The expression levels of p63 and survivin protein were examined by immunohistochemical analysis of 40 cholesteatomas and 5 skin tissues obtained from patients during ear surgery. Results. Expression of p63 protein was diffusely observed in entire samples of cholesteatoma, especially in acquired cholesteatoma, compared with the control group. Congenital cholesteatoma showed variable p63 reactivity in a basal cell-like pattern. Primary and recurrent cholesteatomas showed no significant difference in p63 expression. Survivin was detected in 31 of 40 cholesteatoma samples. Acquired cholesteatomas showed especially increased survivin expression compared with congenital cases. The expression of p63 was correlated with survivin expression.     

Association between human beta defensin expression and cholesteatoma formation

OBJECTIVE: To investigate an association between human beta defensin (hBD) expression and cholesteatoma formation. METHODS: hBD-2 mRNA expressions were assessed in healthy external acoustic meatus skin organ cultures before and after stimulation with Pseudomonas aeruginosa. In addition, hBD-1 and hBD-2 protein production of stimulated and non-stimulated external acoustic meatus skin was visualized by immunohistochemistry. Furthermore, hBD-1 and hBD-2 mRNA expression was analyzed in 25 external acoustic meatus skin, 29 cholesteatoma, and 18 non-cholesteatoma control samples. Non-stimulated meatal tissue preparation did not express hBD-2, whereas incubation with P. aeruginosa demonstrated hBD-2 induction. RESULTS: The hBD-1 mRNA expression was detected in cholesteatoma (14/17), meatal skin, and middle ear mucosa (11/18). hBD-2 mRNA expression was shown in eight cholesteatoma (28.5%) and in three middle ear mucosa tissue samples (37.5%). CONCLUSION: Our data suggest constitutional hBD-1 and inducible hBD-2 expression in chronic middle ear infection and cholesteatoma. Failure of hBD-1 and hBD-2 expression might dispose to exacerbation of cholesteatoma disease. The organ culture model of the external acoustic meatus skin is effective in order to evaluate germ stimulation experiments.     

缺氧诱导因子-1α在中耳胆脂瘤组织中的表达

目的:探讨缺氧诱导因子- 1α(HIF- 1α)在中耳胆脂瘤发病机制中的作用,检测其在中耳胆脂瘤和外 耳道正常上皮中的表达情况。方法:取31例中耳胆脂瘤和10例外耳道正常上皮标本,用免疫组织化学技术检测 HIF-1α蛋白的表达。结果:在中耳胆脂瘤中HIF-1α的表达较外耳道正常上皮明显增多(P<0.05)。结论:中耳 胆脂瘤中HIF-1α的表达显著升高,推测HIF-1在中耳胆脂瘤的发生、发展过程中具有极其重要的作用,并且缺 氧有可能是中耳胆脂瘤发病机制中的一个诱因。     

Cell cycle inhibitory protein p27 in human middle ear cholesteatoma

AIM: To evaluate the immunohistochemical and molecular presentation of protein p27 in cholesteatoma. METHODS: 42 cholesteatoma samples and 6 external ear canal skin (EECS) specimens were investigated and analyzed taking into consideration congenital, acquired, recurrent cholesteatoma, and EECS. RESULTS: The expression of p27 was found in 16 (38.1%) out of 42 specimens of cholesteatoma and in 5 (83.3%) out of 6 specimens of EECS. There was a significant difference in p27-positive staining rate between EECS and cholesteatoma epithelium (p < 0.008). The presence of p27 was detected in 10 cases of acquired cholesteatoma, 2 cases of congenital and 3 cases of recurrent cholesteatoma. There was no significant difference between the presence of p27 in cholesteatoma and EECS (p = 0.01). CONCLUSION: The down-regulation of p27 is a key player in cell cycle control and plays an undefined role in the pathogenesis of all types of cholesteatoma.