PTKs CDK4及p15在中耳胆脂瘤上皮的表达

目的 :从蛋白酪氨酸激酶 (PTKs)信号传递系统及细胞周期调控探讨胆脂瘤上皮细胞增殖的分子机制。方法 :采用免疫组织化学S P方法和计算机图像分析系统 ,观测 30例中耳胆脂瘤上皮及 19例胆脂瘤患者外耳道皮肤中磷酸化PTKs、周期蛋白依赖性激酶 4 (CDK4 )、p15的表达 ,并结合上皮下炎症浸润、骨质破坏程度作统计学分析。结果 :磷酸化PTKs以胞膜表达为主 ,CDK4胞核、胞质均有表达 ,以胞核为主 ,而p15仅以胞核表达。与外耳道皮肤相比 ,胆脂瘤上皮细胞磷酸化PTKs、CDK4、p15表达都显著增强 (P <0 .0 1) ,在重度皮下炎性浸润的上皮细胞磷酸化PTKs、CDK4表达增强 (P <0 .0 1) ;处于高水平磷酸化PTKs表达的胆脂瘤上皮细胞CDK4表达增强 (P <0 .0 1) ;两种骨质破坏程度中上述指标表达的差异无统计学意义 (P >0 .0 5 )。结论 :胆脂瘤上皮细胞具有过度增殖能力 ,同时也存在抑制细胞增殖的机制 ;局部的炎性浸润有增强胆脂瘤上皮细胞增殖能力的倾向     

CDK4、P16在中耳胆脂瘤中的表达及意义

目的：观察细胞周期蛋白依赖性激酶CDK4、细胞周期蛋白依赖性激酶抑制因子P16在中耳胆脂瘤上皮的表达情况,探讨它们在胆脂瘤发病机制中的作用。方法：采用免疫组织化学技术SABC法检测CDK4、P16在中耳胆脂瘤35例、正常外耳道皮肤20例中的表达,并结合胆脂瘤对听小骨骨质破坏程度,采用SPSS11.5软件进行统计学分析。结果：CDK4、P16均表达于胆脂瘤上皮的棘细胞层、颗粒层和角质层,阳性率分别为68.6%、88.6%,高于对照组(前者P=0.011,后者P=0.02)。二者在胆脂瘤上皮的表达有一定的相关性,P＜0.05;与中耳胆脂瘤听小骨骨质破坏程度之间无明显相关性,P＞0.05。结论：CDK4与P16在中耳胆脂瘤发生中是共同而非单独起作用的,从细胞周期的角度可反映胆脂瘤上皮细胞过度增殖的同时也增强负调控因子来抑制增殖,使凋亡同时加快而达到新的平衡。但与骨质破坏程度可能无直接关系。     

中耳胆脂瘤研究进展及其与凋亡抑制蛋白survivin和Bcl-2的关系

国内外学者对中耳胆脂瘤的发病机制做了大量研究，主要集中在上皮过度增殖、骨质破坏吸收和细胞凋亡机制三方面。随着科技的发展，分子生物学技术在研究工作中大量应用，学者们开始从分子生物学角度研究胆脂瘤上皮的特征和演变、骨质破坏、免疫细胞浸润、细胞因子表达与相互作用，把中耳胆脂瘤的病因及发病机制的研究带到了一个新的领域。但是，胆脂瘤的发生、发展有很多复杂的因素，它既是一种炎性疾病，同时又具有肿瘤的某些特性（如上皮细胞过度增殖，侵袭性生长），在其发生、发展中，炎症递质及上皮增生因子（如IL-1、IL-6、IL-8、TNF、TGF、EGF、EGFR等）参与其中，也和某些抑癌基因（如p53、p16、p27等）关系密切。     

胆脂瘤上皮细胞增殖及分子调控机制的免疫组化研究

近年来研究表明胆脂瘤细胞的过度增殖行为与信号传导相关的蛋白酪氨酸激酶(protein tyrosine kinases，PTKs)异常激活有关，信号传导中涉及细胞周期，G1期的重要调控者周期蛋白依赖性激酶4(cyclin dependen kinase4，CDK4)和其特异性抑制因子周期蛋白依赖性激酶抑制物p15调控结果对细胞周期产生重要影响，有决定细胞增殖还是凋亡的作用，因此，我们用超敏免疫组化技术(ultrasensitive immunohistochemistry，SP)检测胆脂瘤上皮细胞磷酸化PTKs、CDK4、p15的表达水平，报道如下。     

胆脂瘤上皮细胞增殖及分子调控机制的免疫组化研究

近年来研究表明胆脂瘤细胞的过度增殖行为与信号传导相关的蛋白酪氨酸激酶(proteintyrosinekinases,PTKs)异常激活有关[1] ,信号传导中涉及细胞周期,G1期的重要调控者周期蛋白依赖性激酶4 (cyclindependenkinase 4 ,CDK4 )和其特异性抑制因子周期蛋白依赖性激酶抑制物p15调控结果对细胞周期产生重要影响,有决定细胞增殖还是凋亡的作用[2 ] ,因此,我们用超敏免疫组化技术(ultrasensitiveimmunohistochemistry,SP)检测胆脂瘤上皮细胞磷酸化PTKs、CDK4、p15的表达水平,报道如下。一、资料与方法1.临床资料:2 0 0 1年8月～2 0 0 2年9月在…     

中耳胆脂瘤上皮中Livin、p73的表达与其发生关系研究

目的 观察凋亡抑制蛋白家族成员Livin及p53基因家族成员p73在中耳胆脂瘤上皮的表达情况,探讨它们在胆脂瘤发病机制中的作用.方法 采用免疫组织化学技术SP法(streptavidin-perosidabe,链霉菌抗生物素蛋白-过氧化物酶连结法)检测30例胆脂瘤标本及16例外耳道正常皮肤组织中的Livin及p73的表达情况,并结合胆脂瘤对听小骨骨质破坏程度,采用SPSS 10.0软件包进行统计学分析.结果 Livin表达于细胞核及部分细胞质,而D73表达于胞浆,上述两指标在中耳胆脂瘤中的阳性表达率分别为76.7%、40%,Livin在胆脂瘤上皮中的表达高于外耳道正常皮肤(P<0.05),而p73在胆脂瘤及外耳道皮肤中的表达差异无显著性(P>0.05),听小骨骨质破坏程度与上述指标的表达差异无统计学意义(P>0.05).结论 Livin在中耳胆脂瘤的异常表达可能在胆脂瘤的发生、发展过程中起相应作用,可能参与胆脂瘤上皮的凋亡调控过程,而p73在中耳胆脂瘤的发生和发展中的作用尚难确定.     

胆脂瘤型中耳炎骨质破坏机制及手术治疗

胆脂瘤型中耳炎是一种是以上皮细胞增生、角化上皮脱屑堆积和骨质破坏为特征的慢性中耳疾病,其骨质破坏作用是导致各种颅内、外并发症的主要原因,胆脂瘤引起骨质破坏的确切机理尚未完全明了,近年来的研究表明与胆脂瘤上皮细胞过度增殖与凋亡、胆脂瘤组织蛋白水解酶、细胞因子及局部炎症等因素有关.有关胆脂瘤型中耳炎手术的术式选择,开放式乳突术腔的处理、内镜技术的应用、咽鼓管功能与鼓室成形术的关系等近年来也倍受关注.     

胆脂瘤型中耳炎发生机制

胆脂瘤型中耳炎是一种常见的慢性中耳炎,其发病机制尚不清楚。本文就胆脂瘤上皮的特性、胆脂瘤骨质破坏机制及胆脂瘤上皮细胞增殖与凋亡的基因调控等方面的研究进展加以综述。     

胆脂瘤型中耳炎发生机制

胆脂瘤型中耳炎是一种常见的慢性中耳炎,其发病机制尚不清楚。本文就胆脂瘤上皮的特性、胆脂瘤骨质破坏机制及胆脂瘤上皮细胞增殖与凋亡的基因调控等方面的研究进展加以综述。     

中耳胆脂瘤分子生物学研究的新进展

中耳胆脂瘤的特征表现为中耳腔内堆积大量的、增殖的、角化的鳞状上皮和邻近骨质的破坏及对听力的影响 ,其侵袭性 ( invasive)、破坏性( aggressive)、迁移性 ( migratory)、非正常调控下的增殖性 ( unrestrained hyperproliferation)、分化变异性 ( altered differenation)、复发性 ( recidivistic)等一直受到耳鼻咽喉科医生的重视 ,本文就最近研究的热点综述如下。1 　 胆脂瘤上皮细胞的增殖与凋亡胆脂瘤具有很强的增殖能力 ,增殖细胞核抗原( PCNA)主要见于细胞合成周期的 S期 ,于各种增殖细胞的胞核有表达 ,可用于衡量细胞的增殖能力。…     

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).     

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。     

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。     

Markers of epidermal proliferation in middle ear cholesteatoma

Middle ear cholesteatoma is characterized by the presence in the middle ear cavity of a stratified squamous epithelium with keratin deposits which by constant proliferation leads to extensive bone destruction. The goal of this study was to evaluate, by immunohistochemical study, the expression of epithelial markers of proliferation--Ki-67 and PCNA in the matrix of cholesteatoma. The materials used in this study were 16 acquired cholesteatoma tissues collected from patients in the age 6-17 years during surgery. The specimens from the skin of the external ear canal were employed as the control. In the immunohistochemical specimens staining intensity and distribution of Ki67 and PCNA positive cells in various layers of the epithelium were assessed in three stages scale. The results were compared to the clinical parameters such as--type of cholesteatoma (pars flaccida or tensa), presence of ear discharge, degree of ossicular destruction and involvement of attic and mastoid. In the cholesteatoma matrix Ki-67 and PCNA positive cells were present in basal and suprabasal cell layers and also more superior layers, unlike the control skin were only basal cells show positive staining. The number of positive cells and intensity of staining was also greater in the cholesteatoma matrix than in skin of external auditory meatus. No correlation was found between results of immunohistochemical examination and clinical parameters.     

A Study on Epidermal Proliferation Ability in Cholesteatoma

With the objective of estimating proliferation ability of epidermis of middle ear cholesteatoma, the difference in proliferating cell nuclear antigen (PCNA) staining between the skin of the bone region of the external ear canal (control skin) and cholesteatoma epidermis and the effects on PCNA staining of subepidermal inflammatory cell infiltration of cholesteatoma were immunohistochemically studied using an antibody against PCNA. Transforming growth factor-α (TGF-α) is known to promote epidermal proliferation based on autocrine mechanism. But it is not clear that cholesteatoma epidermis is actually in the state of hyperproliferation under the effect of this growth factor. To estimate the effect of TGF-α on epidermal proliferation ability, the authors compared the location of PCNA and TGF-α in the same specimen. Unlike the control skin, not only epidermal basal cell layer and suprabasal cell layer, but also more superior layers were found to have high levels of PCNA staining in the epidermis of cholesteatoma. However, in the same cholesteatoma epidermal tissue, the PCNA staining was varied and the difference was ascribable to subepidermal cell inflammation. It appeared that the proliferation ability was high in regions where subepidermal inflammatory cell infiltration was severe. These differences in microenvironment are inferred to greatly affect proliferation ability of cholesteatoma epidermis.     

核转录因子κ-Bp65在中耳胆脂瘤上皮中的表达和意义

目的检测核转录因子-κBp65（nuclear factor kappa Bp65,NF-κBp65）在中耳胆脂瘤上皮中的表达和活化,探讨其在中耳胆脂瘤发病机制中的可能作用。方法采用免疫组织化学SP法检测30例中耳胆脂瘤组织标本与15例正常外耳道皮肤标本中NF-κBp65蛋白的表达。结果NF-κBp65蛋白阳性表达定位于上皮细胞核。NF-κBp65蛋白在中耳胆脂瘤上皮组织中阳性表达率为63.3%,明显高于正常外耳道皮肤组的20.0%（P〈0.01）。结论NF-κBp65蛋白在中耳胆脂瘤上皮的异常表达可能在胆脂瘤的发生、发展过程中起重要作用。胆脂瘤上皮中NF-κBp65的活化可能参与了胆脂瘤上皮细胞过度增殖机制。     

中耳胆脂瘤中热休克蛋白70和细胞周期素依赖蛋白p27的表达及病理意义

目的 探讨热休克蛋白70(heat shock protein 70,Hsp70)和细胞周期素依赖蛋白p~(27)在中耳胆脂瘤中的表达及病理意义。方法 应用免疫组织化学SP法和原位杂交技术,检测40例中耳胆脂瘤上皮中Hsp70、Hsp70mRNA和p~(27)的表达,选用20例正常外耳道皮肤作对照,应用全自动图像分析仪分析各张切片的平均吸光度值。结果Hsp70、Hsp70mRNA和p~(27)在中耳胆脂瘤上皮中表达均明显高于在正常外耳道皮肤中的表达(P<0.05)。而Hsp70和p~(27)二者间在中耳胆脂瘤上皮中表达无相关性(P>0.05)。结论 Hsp70和p~(27)二者可能独自参与了中耳胆脂瘤上皮的增殖、分化及胆脂瘤形成。