基质金属蛋白酶-9和肿瘤坏死因子-α在中耳胆脂瘤中的表达

目的 :观察基质金属蛋白酶 (MMP) 9、肿瘤坏死因子 (TNF) α在胆脂瘤组织中的表达 ,探讨二者在胆脂瘤骨质破坏中的作用。方法 :采用免疫组织化学技术 (SP法 )和计算机图像定量分析法 ,检测TNF α、MMP 9在 2 2例胆脂瘤及 2 0例外耳道皮肤中的表达 ,同时观察二者的表达与临床上听小骨骨质破坏程度的相互关系。结果 :MMP 9表达于胆脂瘤上皮下结缔组织基质细胞 ,TNF α在上皮层及上皮下结缔组织基质细胞中有高度表达 ,定量分析显示胆脂瘤组织中二者的含量显著高于外耳道皮肤 (P <0 .0 5 ) ,并且二者的表达与骨质破坏的程度有关。结论 :MMP 9和TNF α在胆脂瘤组织中呈高表达 ,且与骨质破坏程度呈正比 ,表明其与胆脂瘤的骨质破坏密切相关 ;并且MMP 9和TNF α的表达有相关性 ,提示二者之间可能存在相互作用 ;MMP 9仅在胆脂瘤基质细胞中表达 ,TNF α也在基质细胞中表达 ,提示基质细胞在胆脂瘤的骨质破坏中起着重要的作用     

中耳胆脂瘤基质金属蛋白酶2,9的活性检测及临床意义

目的 :探讨中耳胆脂瘤基质金属蛋白酶 (matrixmetalloproteinases,MMP) 2 ,9的活性水平及临床意义。方法 :应用明胶酶谱法测定 4 1例中耳胆脂瘤和 2 0例外耳道皮肤中MMP2 ,9的活性。结果 :胆脂瘤组织中MMP2和MMP9的平均活性为 (0 .95 4± 0 .4 11)和 (2 .6 76± 0 .734)灰度面积·mg-1·ml-1,而外耳道皮肤的平均活性为 (0 .35 5± 0 .16 0 )和 (1.16 6± 0 .4 4 3)灰度面积·mg-1·ml-1,胆脂瘤组织MMP2和MMP9的活性均较外耳道皮肤高 ,P <0 .0 1,其中MMP9的活性较MMP2高 ,P <0 .0 1,两者呈正相关 ,P <0 .0 1。广泛型胆脂瘤组织中MMP2和MMP9的平均活性为 (1.0 6 2± 0 .4 0 1)和 (2 .94 6± 0 .134)灰度面积·mg-1·ml-1,而局限型胆脂瘤的平均活性为 (0 .70 1± 0 .318)和 (2 .193± 0 .6 0 1)灰度面积·mg-1·ml-1,广泛型胆脂瘤MMP2和MMP9活性较局限型胆脂瘤高 ,P <0 .0 1。复发性与初发性胆脂瘤中MMP2和MMP9活性比较差异均无统计学意义 ,P >0 .0 5。结论 :中耳胆脂瘤中MMP2 ,9活性高于外耳道皮肤 ,且广泛型胆脂瘤高于局限型胆脂瘤 ,提示MMP2 ,9参与胆脂瘤的骨质破坏过程     

MMP-9和EGFR在中耳胆脂瘤中的表达及作用

目的探讨基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)和表皮生长因子受体(epi-dermal growth factor receptor,EGFR)在中耳胆脂瘤骨质破坏中的作用及相互关系。方法采用免疫组织化学技术和计算机图像定量分析法,检测MMP-9、EGFR在22例中耳胆脂瘤上皮组织、10例中耳肉芽组织及22例外耳道深部正常皮肤中的表达,分析二者在中耳胆脂瘤上皮组织的表达与骨质破坏程度的相互关系。结果 MMP-9、EGFR均表达于中耳胆脂瘤上皮组织、中耳肉芽组织及外耳道深部正常皮肤,但中耳胆脂瘤上皮组织中二者的含量显著高于中耳肉芽组织及外耳道深部正常皮肤,MMP-9在中耳胆脂瘤上皮组织中的表达高于EGFR,二者的表达具有相关性,且与骨质破坏的程度有关,骨质破坏范围越广,二者的表达越高(P<0.05)。结论 MMP-9和EGFR在中耳胆脂瘤组织中的表达与中耳胆脂瘤的骨质破坏密切相关,且二者的表达与骨质破坏程度呈正比,二者之间可能存在协同作用;MMP-9在中耳胆脂瘤的骨质破坏中可能具有重要作用。     

基质金属蛋白酶-2及其抑制剂和白细胞介素-6在中耳胆脂瘤的表达及意义

目的研究基质金属蛋白酶-2（matrix metalloproteinase-2,MMP-2）及其抑制剂（tissue inhibitor of metalloproteinase, TIMP-2）和白细胞介素-6（interleukin-6,IL-6）在中耳胆脂瘤中的表达,探讨其表达在胆脂瘤溶骨机制中的作用.方法应用免疫组化技术检测32例患者中耳胆脂瘤组织和10例正常人外耳道皮肤组织中MMP-2、 TIMP-2和IL-6的表达.结果 MMP-2在胆脂瘤上皮各层细胞中均有表达,基底层表达最强,棘层次之,颗粒层及角质层细胞表达较弱;TIMP-2在基底细胞层和棘细胞层有少量表达;IL-6的表达分布于胆脂瘤上皮全层,但在基底层细胞表达最弱,棘细胞层、颗粒层和角质层依次递增,染色最强的部位是颗粒层和角质层,而正常外耳道皮肤未见MMP-2、 TIMP-2、IL-6的表达.结论在胆脂瘤组织中存在MMP-2与TIMP-2的表达失衡,导致胆脂瘤的骨质破坏.同时胆脂瘤上皮高表达IL-6可能是MMPs过表达的原因.     

基质金属蛋白酶在中耳胆脂瘤中的表达

目的研究基质金属蛋白酶MMP(matrix metaIproteinase)2和9在中耳胆脂瘤中的表达情况。方法采用RT-PCR和Western blot方法检测36例中耳胆脂瘤和29例外耳道皮肤组织标本中MMP2和MMP9的基因和蛋白的表达情况。结果 MMP2和MMP9在中耳胆脂瘤中的m RNA水平的表达明显高于外耳道皮肤,差异有统计学意义(P<0.01),MMP2和MMP9在中耳胆脂瘤中蛋白水平的表达高于外耳道正常皮肤,差异有统计学意义(P<0.05)。结论 MMP2、MMP9的高表达可能在中耳胆脂瘤的发生发展过程中起到一定的促进作用,而MMP2和MMP9的高表达可能有助于为中耳胆脂瘤的临床实践提供新的诊疗思路。     

两种基质金属蛋白酶在胆脂瘤和中耳癌中的表达

目的探讨基质金属蛋白酶（matrix metalloproteinases, MMP）的MMP-2和MMP-9在胆脂瘤和中耳癌中的表达差异及其在骨浸润作用。方法采用免疫组织化学方法检测36例胆脂瘤上皮,10例胆脂瘤外耳道皮肤、16例中耳癌组织及正常组织中MMP-2和MMP-9的表达。结果①MMP-2和MMP-9在胆脂瘤上皮染色最强。而且在上皮组织的各层细胞均有表达,基底膜染色最强。②胆脂瘤上皮中的MMP-2和MMP-9的表达明显高于其对应的外耳道皮肤和正常外耳道上皮。MMP-2阳性表达在胆脂瘤上皮中为83%,胆脂瘤外耳道上皮为50%,正常外耳道皮肤2例有微弱表达。③16例中耳鳞癌组织中,MMP-9的阳性表达率为50%（8/16）,MMP-2的阳性表达率为56.25%（9/16）,表达水平与肿瘤的病理分级和临床分期有关。④中耳癌组织中MMP-2的表达与MMP-9表达不一致,而胆脂瘤组织中MMP-2和MMP-9的表达密切相关。结论胆脂瘤向周围骨质侵袭的特性与中耳癌相似,MMPs家族及其抑制剂之间的平衡紊乱所致的蛋白水解酶活性增高是引起中耳胆脂瘤骨吸收的原因之一。     

MMP-9在中耳胆脂瘤中的表达和意义

目的 研究基质金属蛋白酶-9(matrix metalloproteinase,MMP-9)在中耳胆脂瘤上皮中的表达,并探讨其与中耳胆脂瘤的血管形成以及胆脂瘤侵蚀行为的关系.方法采用免疫组织化学SP法检测MMP-9、Ⅳ型胶原蛋白在44例患者中耳胆脂瘤组织标本与10例正常外耳道组织中的表达.结果 MMP-9在胆脂瘤上皮各层均有表达,在外耳道皮肤中为弱阳性表达;胆脂瘤基质周围微血管计数均数高于正常外耳道皮肤,二者差异具有统计学意义(P＜0.05);MMP-9在胆脂瘤中的表达与周围微血管计数间呈正相关关系.结论 MMP-9可能在中耳胆脂瘤血管生成中起重要作用.     

MMP-13和MMP-9与中耳胆脂瘤组织侵袭性的研究

目的究MMP-13和MMP-9在中耳胆脂瘤组织的表达及其临床意义,探讨其相互作用机制及骨质破坏的相关性。方法用免疫组化SP法检测176例中耳胆脂瘤及外耳道正常皮肤中MMP-13和MMP-9的表达,并进行平均光密度值的定量分析与周围骨质破坏范围情况。结果 MMP-13和MMP-9均表达于胞质中,MMP-13和MMP-9在胆脂瘤组织中均呈高表达,平均光密度值显著高于外耳道正常上皮组织,且两者表达呈正相关(P0.01)。随着周围骨质破坏范围增大,MMP-13和MMP-9表达越高。结论 MMP-13和MMP-9共同参与胆脂瘤组织对骨质的破坏,可能主要通过MMP-13激活MMP-9途径来参与胆脂瘤组织的侵袭。     

两种基质金属蛋白酶在胆脂瘤和中耳癌中的表达

目的 探讨基质金属蛋白酶（matrix metalloproteinases,MMP)的MMP－2和MMP－9的胆脂瘤和中耳癌中的表达差异及其在骨浸润作用。方法 采用免疫组织化学方法检测36例胆脂瘤上皮，10例胆脂瘤外耳道皮肤、16例中耳癌组织及正常组织中MMP－2和MMP－9的表达。结果 ①MMP－2和MMP－9在胆脂瘤上皮染色最强。而且在上皮组织的各层细胞均有表达，基底膜染色最强。②胆脂瘤上皮中的MMP－2和MMP－9的表达明显高于其对应的外耳道皮肤和正常外耳道上皮。MMP－2阳性表达在胆脂瘤上皮中为83％，胆脂瘤外耳道上皮为50％，正常外耳道皮肤2例有微弱表达。③16例中耳鳞癌组织中，MMP－9的阳性达率为50％（8／16），MMP－2的阳性表达率为56.25%(9/16)，表达水平与肿瘤的病理分级和临床分期有关。④中耳癌组织中MMP－2的表达与MMP－9表达不致，而胆脂瘤组织中MMP－2和MMP－9的表达密切相关。结论 胆脂瘤向周围骨质侵袭的特性与中耳癌相似，MMPs家族及其抑制剂之间的平衡紊乱所致的蛋白水解酶活性增高是引起中耳胆脂瘤骨吸收的原因之一。     

MMP9及VEGF诱导血管生成在中耳胆脂瘤增殖与侵袭中的作用

目的探讨基质金属蛋白酶(matrix metalloproteinases,MMP)9及血管内皮生长因子(vascular endothelial growth factor,VEGF)诱导血管生成在中耳胆脂瘤增殖与侵袭中的作用。方法通过免疫组化方法检测MMP9、VEGF、增殖细胞核抗原(Ki-67)和CD105在45例中耳胆脂瘤组织和10例正常外耳道皮肤中的表达。结果 (1)中耳胆脂瘤上皮下结缔组织中平均微血管密度(MVD)高于正常外耳道皮肤,差异有统计学意义(t=8.905,p<0.001);(2)随着胆脂瘤上皮下结缔组织MVD的增加,其Ki-67阳性表达率也相应地增加;(3)中耳胆脂瘤上皮中VEGF和MMP9的平均光密度值高于正常外耳道皮肤,差异有统计学意义(t=4.804,p<0.01;t=3.184,p<0.01);随着MMP9和VEGF平均光密度值增加,胆脂瘤上皮下结缔组织微血管密度(MVD)也相应地增加;(4)相关性分析显示VEGF和MMP9在诱导血管生成过程中具有协同作用(r=0.851,p=0.000)。结论 (1)中耳胆脂瘤上皮下结缔组织中存在微血管增生。(2)胆脂瘤上皮下结缔组织中微血管增生可以为胆脂瘤上皮的异常高度增殖与侵袭提供较丰富的血供和营养。(3)MMP9和VEGF在诱导胆脂瘤血管生成过程中具有协同作用。     

Expression of matrix metalloproteinasa-2,9 in cholesteatoma and middle ear cancer]

OBJECTIVE: To study various expressions and roles of MMP-2, 9 in cholesteatoma and middle ear cancer. METHODS: MMP-2 and MMP-9 were detected with immunohistochemical methods using monoclonal antibody in 36 cholesteatomas, 10 CAMS and 16 middle ear cancer. RESULTS: The stain in cholesteatoma was stronger than that in CAMS and NAMS. Expressions of MMP-2, 9 were seen in all layers of the epithelium in CA and the stain was strongest in basement membrane. The positive rates of stain was 83% and 50% in CA and CAMS respectively. There was weak stain in 2 NAMS. In middle ear cancer, the expression rates of MMP-9 and MMP-2 were 50% (8/16) and 56.25% (9/16) respectively. The expression level was correlated with tumor differentiation and its clinical stage. Expressions of MMP-2 and MMP-9 were closely related in cholesteatoma. There was no relationship between expressions of MMP-2 and MMP-9 in middle ear cancer. CONCLUSION: Cholesteatoma has a similar feature with middle ear cancer in invading surrounding of surround bone matrix. The disorder between MMPs and its inhibitor was one of reasons for bone absorption of cholesteatoma and middle ear cancer.     

Stromal expression of matrix metalloprotease-9 in middle ear cholesteatomas

BACKGROUND: Matrix metalloprotease-9 (MMP-9) plays an important role in the degradation of extracellular matrix (ECM). In cholesteatoma there are significant changes in the composition of the ECM, supporting the view of a disturbed cell-ECM interaction. It has been reported that in cholesteatoma tissues, MMP-9 is expressed in epithelial cells but not in stromal cells. In this study, we report conflicting results regarding the identity of the cholesteatoma cells that express MMP-9. METHODS: An immunohistochemical assay was performed to examine the expression of MMP-9 in 25 cases (formalin-fixed paraffin-embedded sections) of cholesteatoma, using an MMP-9-specific monoclonal antibody. RESULTS: In all cholesteatomas examined, MMP-9 was expressed in the stromal cells but not in the epithelial cells. MMP-9 immunostaining was undetectable in external auditory meatal skin (n = 5) and tympanic membrane (n = 3). CONCLUSIONS: These results suggest that the MMP-9 expressed in stromal cells may be involved in the disturbed cell-ECM interaction associated with the process of cholesteatoma formation.     

Die experimentelle Erzeugung von Cholesteatomen

Summary In 45 rabbits, Marbagelan (a gelfoam like substance) soaked with Tris-HCl-buffer and histoacryl was applied on the meatal epithelium near the tympanic membrane. In 87% of the ears a cholesteatoma of different localisation (external auditory canal, lateral attic wall, ear drum middle ear cleft) developped. In 20 cases (34%) cholesteatoma appeared within four weeks. Continuous irritation is probably responsible for cholesteatoma formation.     

Expression of a cell-cycle-associated nuclear antigen (MIB 1) in cholesteatoma and auditory meatal skin

Middle ear cholesteatoma is often invasive with consequent bone destruction. Inflammatory stimulation of the underlying connective tissue, as well as an autocrine mechanism, may be responsible for the dys-regulation and abnormal proliferative features of the keratinocytes in cholesteatoma. Comparative investigations were performed to assess the epithelial cell kinetics of cholesteatoma and normal auditory meatal skin. Monoclonal antibody MIB 1 immuno-staining (which recognizes a nuclear antigen expressed by dividing cells) was applied using the alkaline phosphatase antialkaline phosphatase im-munolabeling method. Specimens of normal auditory meatal skin (n = 7) revealed an average MIB 1 score (quotient of the MIB 1-positive cells and the total number of cells) of 7.6 ± 2.2%. Cholesteatoma samples (n = 13) showed an average MIB 1 score of 17.4 ± 8.9% and a heterogeneity of proliferating epithelial areas. Epithelial cones growing toward the underlying stroma exhibited high mitotic activity. Statistically, the results of this study confirm a highly significant increase in the proliferation rate of cholesteatoma keratinocytes, which had an MIB 1 score that was 2.3 times higher than the score for keratinocytes of normal external auditory meatal skin.     

Matrixmetalloproteinase-1 in cholesteatoma,middle ear granulations and deep meatal skin: a comparative analysis

Nine cholesteatomas, seven middle ear granulations and five deep meatal skin specimens were analysed for gelatinase activity at molecular weights corresponding to those of Matrix metalloproteinase-1 (MMP-1) using SDS PAGE Zymography. Gelatinase activity at 41–43 kDa and 45–47 kDa was investigated. Western blotting was employed using a primary monoclonal antibody to MMP-1 to provide a qualitative assessment of MMP-1. Western blotting was also used with a monoclonal antibody to MMP-3 to discover if MMP-3 gelatinase activity occurring around the molecular weight of MMP-1 may have contributed to the results. A significantly higher expression of activity was recorded in cholesteatoma and middle ear granulations at 45–47 kDa in comparison with deep meatal skin. Western blotting indices were to be present in all of the cholesteatoma specimens tested. Only one of the specimens (cholesteatoma) tested showed any MMP-3 presence.     

Matrix metalloproteinase-2 and Matrix metalloproteinase-9 in cholesteatoma and deep meatal skin

A qualitative and quantitative study of the presence of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9) in cholesteatoma was performed. Ten cholesteatoma and four deep meatal skin specimens were analysed for gelatinase activity at molecular weights corresponding to MMP-2 and MMP-9 using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE) Zymography. Gelatinase activity at 72 kDa and 92 kDa was investigated. Western blotting was employed using primary monoclonal antibodies to provide a qualitative assessment of MMP-2 and MMP-9. Non-parametric data analysis using the Mann–Whitney U test did not show a significant difference in expression of MMP-2 (P = 0.51) or MMP-9 (P = 0.14) between the two tissue types. Western blotting showed the presence of both MMP-2 and MMP-9 in the majority of specimens, both cholesteatoma and deep meatal skin.     

The localization of matrix metalloproteinases-8 and -13 in cholesteatoma, deep-meatal and post-auricular skin: a comparative analysis

CONCLUSION. The presence of matrix metalloproteinase (MMP)-8 and MMP-13 was found to be significantly higher in cholesteatoma compared with post-auricular skin. The results show that the control group used has implications for further studies. OBJECTIVES. To compare the presence of MMP-8 and MMP-13 in cholesteatoma, deep meatal and post-auricular skin. Our null hypothesis was that there was no difference in expressions of MMP-8 and MMP-13 in the three groups. MATERIALS AND METHODS. The study was carried out in a secondary care specialist centre and used prospective retrieval of specimens for immunohistological localization of MMP-8 and MMP-13. Eleven patients undergoing cholesteatoma surgery were recruited for the study. Eleven cholesteatoma specimens, 10 deep meatal skin specimens and 10 post-auricular skin specimens were analysed. Specimens were analysed by immunohistochemistry using monoclonal antibodies to MMP-8 and MMP-13. Two observers scored the slides independently in a blind fashion. RESULTS. The presence of MMP-8 and MMP-13 was found to be significantly higher in cholesteatoma compared to post-auricular skin (p=0.02, p=0.03, respectively). There were no significant differences in expression of MMP-8 and MMP-13 between cholesteatoma and deep meatal skin (p=0.08, p=0.09, respectively). There were no significant differences in the control groups.     

RANKL和IL-17A在中耳胆脂瘤中的表达及相关性研究

目的观察破骨细胞分化因子（RANKL）和白介素-17A（IL-17A）在中耳胆脂瘤组织中的表达,探讨二者在中耳胆脂瘤骨质破坏机制中的作用。方法采用免疫组织化学两步法和计算机图像定量分析法,检测RAN-KL、IL-17A在25例中耳胆脂瘤上皮、16例胆脂瘤周围肉芽组织及15例正常外耳道皮肤中的表达,同时观察二者的表达与临床听小骨骨质破坏程度的相互关系。结果（1）RANKL、IL-17A在胆脂瘤上皮及胆脂瘤周围肉芽组织中表达增高,分别与正常对照组比较,差异均有统计学意义（tRANKL=9.864、5.630,均P〈0.05;tIL-17A=13.905、9.011,均P〈0.05）,且二者的表达与骨质破坏程度相关。（2）在胆脂瘤上皮及胆脂瘤周围肉芽组织中,RANKL与IL-17A的表达呈正相关（r上皮=0.692,r肉芽=0.538,P〈0.05）。结论RANKL及IL-17A在中耳胆脂瘤组织中呈高表达,与骨质破坏程度成正比,表明其与胆脂瘤骨质破坏机制密切相关;RANKL与IL-17A表达有相关性,提示二者之间可能存在相互作用。