猴病毒40T抗原和P53蛋白在毛丝鼠中耳上皮细胞系的表达及其意义

目的　证实猴病毒 40 (simianvirus40 ,SV40 )T抗原 (largeT antigen ,TAg)和P5 3蛋白在毛丝鼠 (chinchilla)中耳上皮细胞系细胞核内的表达 ,初步探讨它们在细胞无限增殖 (immortalization)过程中的作用。方法　应用免疫荧光组织化学原理 ,分别检测SV40TAg和P5 3蛋白在培养液内注入腺病毒 12 猴病毒 40混合病毒 (实验组 )及未在培养液内注入上述病毒 (对照组 )的体外培养毛丝鼠中耳上皮细胞核内的表达。结果　实验组的 8个培养皿中 ,1个培养皿内的细胞核内表达SV40TAg和P5 3蛋白 ,且已持续培养 46代以上 ,形成细胞系 ;其余 7个培养皿内的细胞和对照组细胞核内均不表达SV40TAg和P5 3蛋白 ,且培养到 6～ 8代时均自行凋亡。结论　SV40TAg和P5 3蛋白在细胞无限增殖过程中起重要的作用 ;它们在细胞核内的表达对毛丝鼠中耳上皮细胞系的建立可能具有密切的相关性。     

喉鳞癌中增殖细胞核抗原和p53蛋白表达的临床意义及其…

本文以ＬＳＡＢ免疫组化方法，用单克隆抗体ＰＣ１０和ＤＯ－１对３０例喉鳞癌及１０例对照组（声带息肉）的增殖细胞核抗原（ＰＣＮＡ）与ｐ５３蛋白表达进行研究，探讨其生物学意义。结果表明；１）喉鳞癌组织ＰＣＮＡ阳性表达率为１００％（ＰＣＮＡ－Ｌｌｓ１６％～９２％），强阳性率为３６．７％（１１／３０），对照组６例阴性，４例弱阳性表达（ＰＣＮＡ－ＬＩｓ〈１０％），试验组阳性表达率及表达程度明显高于对照组（Ｐ〉     

P53蛋白及增殖细胞核抗原（PCNA）在喉癌细胞中的表达及意义

采用ＡＢＣ免疫组化法对７８例喉癌行回顾性及前瞻性研究，测定喉癌细胞Ｐ５３蛋白及增殖细胞核抗原的表达，结果临床病理及随访资料对Ｐ５３蛋白过度表达在喉癌发生发展中的作用及意义进行探讨。     

喉癌微血管生成中P53nm23蛋白和血管内皮生长因 …

目的 探讨Ｐ５３、ｎｍ２３蛋白和血管内皮生长因子（ｖａｓｃｕｌａｒ ｅｎｄｏｔｈｅｌｉａｌ ｇｒｏｗｔｈ ｆａｃｏｒ,ＶＥＧＦ）在喉癌微血管生成及转移中的作用。方法 通过免疫组化ＳＰ法对４２例喉癌标本中Ｐ５３、ｎｍ２３蛋白、ＶＥＧＦ及微血管密度（ｍｉｃｒｏｖｅｓｓｅｌ ｄｅｎｓｉｔｙ,ＭＶＤ）进行了检测。结果 喉癌中Ｐ５３、ｎｍ２３蛋白及ＶＥＧＦ的阳性表达率分别占４７．６％、５７１％和７１．４％。     

喉癌组织及手术切缘P53和P16蛋白表达与局部复发的关系

目的 用免疫组织化学方法监测Ｐ１６和Ｐ５３蛋白在喉癌手术切缘中表达情况及其与肿瘤局部复发的关系。方法 选择ＨＥ染色病理学检查手术切缘为阴性但术后肿瘤局部复发的５２例喉癌作为复发组,并按照患才的年龄、性别、临床分型、ＴＮＭ分期、病理分级、术前放疗剂量等可能影响因素选择４８例随访５年以上无肿瘤局部复发者为无复发组。对２组手术切缘和肿瘤标本同时行Ｐ１６和Ｐ５３蛋白 免疫组织化学检查。结果 在１００例喉癌组织中Ｐ５３蛋白的表达率和Ｐ１６蛋白的丢失率分别为５４％和４０％,喉癌组织中Ｐ１６和Ｐ５３蛋白表达同时异常的喉癌患者当其手术切缘粘膜中Ｐ１６和Ｐ５３蛋白表达也同时异常时,其术后肿瘤局部复发率为８５％。Ｐ１６和Ｐ５３蛋白表达与喉癌的临床分型和Ｔ分期无关。结论 采用免疫组化方法联合检测Ｐ１６和Ｐ５３蛋白在喉癌组织和手术切缘     

口腔粘膜鳞癌组织中P53和nm23蛋白的表达分析

目的：研究口腔粘膜鳞癌组织中P53和nm23蛋白的表达及意义，方法：采用免疫组化方法检测100例口腔粘膜鳞癌组织P53和nm23蛋白的表达状况。结果：口腔粘膜鳞癌组织中P53蛋白的阳性表达率为58．0，nm23的蛋白的阳性表达率为40．0％（P＜0．05），颈淋巴结有转移者较无转移者P^53蛋白阳性表达率高，而nm23蛋白阳性表达率低，结论：P53和nm23蛋白表达状况与口腔粘膜鳞癌颈淋巴结转移有关。     

nm23-H1蛋白和P53蛋白在鼻咽癌的表达

为观察ｎｍ２３－Ｈ１基因产物和Ｐ５３基因产物在鼻咽癌（ＮＰＣ０的表达状态，春一ＮＰＣ的发生及转移、预后的产生。采用Ｓ－Ｐ免疫组织化学方法检测ｎｍ２３－Ｈ１基因产物和Ｐ５３基因产物的表达。结果ｎｍ２３－Ｈ１蛋白的表达率为５０．４％，Ｐ５３蛋白表达率为９０．４％。ｎｍ２３－Ｈ１蛋白在颈部有淋巴结转移包块才的表达率为２７．９％，低于无颈部淋结转移包块者的表达率，两者有非常显著性差异。ｎｍ２３－Ｈ１蛋白及     

喉鳞癌中增殖细胞核抗原和p53蛋白表达的临床意义及其相互关系

本文以ＬＳＡＢ免疫组化方法，用单克隆抗体ＰＣ１０和ＤＯ－１对３０例喉鳞癌及１０例对照组（声带息肉）的增殖细胞核抗原（ＰＣＮＡ）与ｐ５３蛋白表达进行研究，探讨其生物学意义。结果发现：１）喉鳞癌组织ＰＣＮＡ阳性表达率为１００％（ＰＣＮＡ－ＬＩｓ１６％～９２％），强阳性率为３６７％（１１／３０），对照组６例阴性，４例弱阳性表达（ＰＣＮＡ－ＬＩｓ＜１０％），试验组阳性表达率及表达程度明显高于对照组（Ｐ＜００５）。２）ｐ５３蛋白在喉鳞癌中阳性表达率为６６７％，对照组全部阴性，有显著差异（Ｐ＜００５）。３）在喉鳞癌组织中，ＰＣＮＡ和ｐ５３蛋白表达与临床分型、Ｔ分期及病理分化程度无显著关系（Ｐ＞００５），与淋巴转移及术后生存率关系密切（Ｐ＜００５）。４）ＰＣＮＡ与ｐ５３蛋白表达正相关。提示：１）喉鳞癌的增殖活性显著增强，ｐ５３基因突变与喉鳞癌的发生和发展有关。２）检测ＰＣＮＡ和ｐ５３蛋白有助于喉鳞癌的早期诊断和鉴别诊断，帮助治疗决策及判断预后。３）ｐ５３对ＰＣＮＡ有调节作用，为研究肿瘤的发生机理和寻找防治对策提供了新的理论依据。     

Immortalization of chinchilla middle ear epithelial cells by adenovirus 12-simian virus 40 hybrid virus.

In order to study the cellular and molecular mechanisms of the pathogenesis of otitis media, a chinchilla middle ear epithelial cell line (CMEE-1) with differentiated cell characteristics was established by infection of a primary culture with the adenovirus 12-simian virus 40 (Ad12-SV40) hybrid. This cell line has been in continuous culture for 42 passages, whereas the parent cells underwent senescence and died at the 8th passage. The cell line also retains epithelial morphology and expresses cytokeratin polypeptides 4, 7, and 18, characteristic markers for epithelia. In Western blots of cell proteins, bands at 94 and 53 kd were labeled after binding antibodies against SV40 large T antigen and p53, respectively. Karyotype analysis showed that the cell line is derived from chinchilla epithelial cells. These findings confirm that the cell line is a chinchilla epithelial cell immortalized by the hybrid virus.     

p53 Expression and keratinocyte hyperproliferation in middle ear cholesteatoma

Keratinocytes in middle ear cholesteatoma have hyperproliferative properties. There is controversy regarding the role of p53 and its effect on cellular proliferation in cholesteatoma. This study was instituted to examine this. Cholesteatoma and deep meatal skin control specimens were analysed for MIB-1 (n = 7, controls = 7), a marker of cellular proliferation, and p53 (n = 17, controls = 17) expression by immunocytochemistry. Expression of p53 was minimal or absent in both cholesteatoma and controls (P = 0.2). MIB-1 expression was higher, but not significantly so, in cholesteatoma than in controls (P = 0.09). Our study has shown no significant p53 expression in cholesteatoma epithelium. This suggests that there is no dysfunction in the p53-mediated cell cycle control mechanisms in cholesteatoma.     

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.     

Biochemical characterization of mucous glycoproteins secreted by in vitro chinchilla middle ear epithelial cells

Summary A method for middle ear epithelial (CMEE) cell culture with active mucus secretory function has been successfully developed, using the chinchilla as an animal model. CMEE cells were dissociated by protease digestion from the middle ear mucosa. The CMEE cells grown in primary culture incorporated [³H] glucosamine into a glycoconjugate after its release into medium. This substance was characterized biochemically as mucin, although the production of mucin by the cells required growth on a substratum of collagen gel. These cultures provide an excellent model for studying factors that regulate synthesis and secretion of glycoproteins in CMEE cells.     

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。     

Immortalization of rat middle ear epithelial cells by adeno 12-SV40 hybrid virus

Rat middle ear epithelial cells were infected with the adeno 12-SV40 hybrid virus. The cell line thus obtained displays features of primary cultured epithelial cells in both light microscopic and ultrastructural examinations. The immortalized cells have been in continuous proliferation for 40 passages and more than 17 months. Immunohistochemical analysis of the immortalized cells was positive for the SV40 T antigen and the tumor suppressor protein p53. The cells also stained positive for cytokeratin, an epithelial cell marker, and negative for vimentin, a fibroblast marker. These results, together with karyotype analysis, indicate that this cell line originated from rat middle ear epithelial cells and retains the characteristics of epithelial cells. This cell line will be useful for studying the normal cellular biology of middle ear epithelial cells, as well as the cellular and molecular mechanisms involved in the bacteria-middle ear epithelial cell interaction.     

血管内皮生长因子在中耳胆脂瘤中的表达及其临床意义

目的本研究的主要目的是分析与正常中耳黏膜和外耳道皮肤相比血管内皮生长因子(VEGF)在中耳胆脂瘤中的定位和分布.探讨VEGF与中耳胆脂瘤的血管形成以及胆脂瘤侵蚀性行为的关系.方法使用免疫组织化学的方法研究VEGF的表达;使用自动影像分析系统计算所形成血管的数目(Ⅳ型胶原蛋白染色).结果胆脂瘤上皮层基底部角质细胞可见VEGF的免疫反应活性呈强阳性.胆脂瘤基质周围与正常中耳黏膜上皮下结缔组织和正常外耳道皮肤相比,VEGF的免疫反应活性有显著性差异.随着胆脂瘤基质周围炎症程度的增加,其微血管计数的均数也相应地增加,由9.3±3.6血管数/mm2(炎症Ⅰ级)增加到21.1±3.4血管数/mm2(炎症Ⅱ级)和32.5±9.5血管数/mm2(炎症Ⅲ级)(p＜0.05).结论VEGF诱导的新生血管形成能够使角质细胞迁移进入中耳腔.因此,新生血管形成是中耳胆脂瘤侵蚀性行为的一个关键性的因素.