Survivin、Ki67在牙源性囊肿上皮组织中的表达及意义

目的：探讨根端囊肿、含牙囊肿及角化囊肿衬里上皮中Survivin、Ki67的表达及意义。方法：采用免疫组化法检测12例根端囊肿、20例含牙囊肿、22例角化囊肿中Survivin、Ki67表达水平,并加以分析。结果：Survivin在根瑞囊肿中无表达,在含牙囊肿和角化囊肿中表达的阳性率分别为10％、63．6％,角化囊肿OD值显著高于根端囊肿和含牙囊肿（P〈0．05）;Ki67在3种组织中均有表达,角化囊肿中Ki67-ＬＩ显著高于根端囊肿和含牙囊肿（Ｐ〈0．05）;角化囊肿中Survivin阳性表达的Ki67-LI显著高于表达阴性者（Ｐ〈0．05）,且Survivin表达与Ki67表达呈正相关（r=0．452,P〈0．05）。结论：survivin、Ki67在牙源性囊肿中的表达差异显示了它们具有不同的增殖和分化过程。     

细胞角蛋白及其mRNA在根端囊肿上皮衬里中的表达

目的本研究目的是探讨根端囊肿上皮衬里细胞角蛋白及其mRNA的表达情况.方法检测52例根端囊肿衬里上皮的细胞角蛋白(CK8,CK13和CK18)的表达,其中采用免疫组化染色法检测32例上颌根端囊肿和20例下颌根端囊肿;采用原位杂交法检测24例上颌根端囊肿和13例下颌根端囊肿CK-mRNA表达;采用逆转录聚合酶链反应(RT-PCR)法检测24例上颌囊肿.结果上颌根端囊肿中CK8,CK13,CK18阳性的鳞状上皮衬里分别是20例,29例和19例,下颌根端囊肿中表达阳性的分别为10例,20例和11例.上颌根端囊肿中[CK18( )-CK13(-)]3例,[CK18( )-CK13( )]13例,[CK18(-)-CK13( )]13例,而下颌根端囊肿中[CK18( )-CK13(-)]0例,[CK18( )-CK13( )]11例,[CK18(-)-CK13( )]9例.原位杂交显示24例上颌根端囊肿和13例下颌根端囊肿分别有9例和4例囊肿衬里中有CK18-mRNA表达.RT-PCR检测发现CK18-mRNA和CK13-mRNA在正常鼻窦粘膜和牙龈上皮(对照组)及上颌囊肿衬里都有表达.结论CK18-mRNA和CK13-mRNA在根端囊肿鳞状上皮和柱状上皮基本上都有表达.上颌根端囊肿中CK蛋白及其CK18-mRNA表达的不同,可能是囊肿上皮衬里基因型发生转变的结果.     

根侧角化囊肿的CK18及基因表达的研究

目的:对根侧角化囊肿与其它角化囊肿衬里上皮细胞角蛋白18(CK18)基因表达是否一致进行探讨。方法:1例根侧角化囊肿、2例颌骨牙源性角化囊肿和1例含牙囊肿的衬里上皮,使用CK18基因探针进行原位杂交,检测CK18 mRNA在衬里上皮细胞层的原位表达;同时进行CK18、PCNA、p53、p21、Bcl-2、Fas/Fas-L、Bax(B-9)、WAF-1、CEA的单克隆抗体免疫组织化学染色。结果:原位杂交法显示CK18 mRNA在根侧角化囊肿的表达为强阳性,其它角化囊肿衬里上皮表现为弱阳性;免疫组化染色结果显示PCNA和Bcl-2在根侧角化囊肿中表达为强阳性,在2例颌骨牙源性角化囊肿表达为弱阳性,在含牙囊肿中有表达;而p53、p21、Fas/Fas-L、CEA在病例中均为阴性表达;Bax(B-9)、WAF-1在1例角化囊肿中表达为弱阳性,在其余3例中均为阴性表达。结论:根侧角化囊肿与其他部位角化囊肿相比较,强阳性表达反映增殖能力的相关蛋白及阴性表达与凋亡有关蛋白、抑癌基因,提示根侧角化囊肿可能比其它部位角化囊肿增殖或复发的潜力更高。     

牙源性角化囊肿中Survivin的表达及其与Caspase-3和p53的关系

目的:初步探讨凋亡相关蛋白Survivin、Caspase-3和p53在牙源性角化囊肿(odontogenic keratocyst,OKC)中的表达及关系。方法:TUNEL法检测40例OKC(原发20例,复发20例)中凋亡细胞的分布;免疫组织化学方法检测OKC中Survivin、Caspase-3和p53蛋白的表达。结果:OKC中凋亡细胞见于衬里上皮表层细胞;Survivin蛋白阳性染色见于26例(65%)OKC上皮基底层及基底上层细胞中,Caspase-3蛋白阳性染色见于18例(45%)OKC上皮表层细胞中,p53蛋白在OKC中不表达。结论:凋亡相关蛋白Survivin和Caspase-3在OKC形成和发展中发挥一定作用。     

牙源性囊肿抗增殖细胞核抗原抗体细胞动态分析

作者应用抗增殖细胞核抗原(PCNA)抗体免疫组织化学方法,研究囊肿壁内衬表皮细胞动态。以牙源性囊肿手术摘除的囊肿壁内衬表皮完整者作为标本。其中根端囊肿和含牙囊肿各20例,始基囊肿及牙源性角化囊肿25例,钙化性囊肿1例。造釉细胞瘤15例和正常牙龈5例作对照。 结果:囊肿内衬上皮及结缔组织的细胞核PCNA略呈阳性,尤其在囊肿衬里上皮的基底细胞及其上的2～3层细胞多见。根端囊肿表现在组织学上炎症反应     

CK18和CK13在含有纤毛柱状细胞的根端囊肿衬里上皮中的表达

目的:探讨细胞角蛋白CK13和CK18在含有纤毛柱状细胞的根端囊肿衬里上皮中的表达意义。方法:利用单克隆CK18和CK13抗体,采用免疫组化SP法检测32例含有纤毛柱状细胞的根端囊肿衬里上皮中CK13和CK18的表达情况。结果:CK18在此32例含纤毛柱状上皮细胞的根端囊肿衬里中呈现3种表达形式:①伴随纤毛柱状上皮细胞呈连续带状表达在根端囊肿衬里上皮的表层;②呈片段状表达在根端囊肿衬里上皮中;③呈零星散在状表达于衬里上皮细胞中。CK13表达在根端囊肿衬里上皮的鳞状细胞中。一部分基底细胞层细胞和上皮片断既不表达CK18,也不表达CK13。结论:根端囊肿衬里上皮中除含有鳞状细胞外还含有以多种形式分布存在的纤毛柱状细胞及可能处于化生过渡阶段的细胞或上皮发生细胞。根端囊肿衬里上皮中存在着多种细胞类型为根端囊肿的发生机制及组织来源的进一步研究探讨提供了一定的依据。     

牙源性角化囊肿细胞增殖抗原和表皮生长因子受体表达

目的　探讨牙源性角化囊肿衬里上皮细胞的增殖特点。方法　采用免疫组化染色方法 ,对牙源性角化囊肿、成釉细胞瘤、含牙囊肿、正常口腔粘膜上皮中细胞增殖抗原 Ki- 6 7和表皮生长因子受体 (EGFR)的表达进行分析比较。结果　牙源性角化囊肿中 Ki- 6 7表达较含牙囊肿高 ,与正常口腔上皮相似 ;复发的与未复发的牙源性角化囊肿 Ki- 6 7指数无显著性差异。牙源性角化囊肿中 EGFR表达呈阳性。结论　牙源性角化囊肿上皮增殖活跃 ,上皮增殖生长可能与表皮生长因子家族有关。     

牙源性囊肿上皮增殖活性的研究

目的：研究牙源性角化囊肿,含牙囊肿,根尖囊肿3种主要的牙源性囊肿衬里上皮的细胞增殖活性,方法：应用Ki-67单克隆抗体免疫组化LSAB法对30例牙源性囊肿进行免疫组化染色,结果通过计算机图像分析,计算单位面积衬里上皮内（mm2)阳性细胞数,进行统计学分析,结果：牙源性角化囊肿衬里上皮有较多的Ki-67阳性细胞,明显高于含牙囊肿和根尖囊肿;正常口腔粘膜未见Ki-67阳性表达,结论：Ki-67在不同的牙源性囊肿中表达的差异显示了它们具有不同的增殖和分化过程。     

生存素、半胱氨酸天冬氨酸蛋白酶3在口腔癌前病变及口腔癌中的表达

目的 探讨生存素(survivin)、半胱氨酸天冬氨酸蛋白酶3(caspase-3)在口腔癌发生中的作用,以期为口腔癌的监测和治疗提供参考.方法 选取首都医科大学口腔医学院病理科存档石蜡标本45例,其中口腔白斑伴上皮轻-中度异常增生16例,口腔白斑伴上皮重度异常增生12例,口腔高-中分化鳞状细胞癌17例;正常口腔黏膜组织10例.采用免疫组化SP法对生存素、caspase-3的表达进行检测;脱氧核糖核苷酸末端转移酶介导的原位缺口末端标记法对细胞凋亡指数进行检测.结果 生存素表达在正常黏膜、白斑伴上皮轻-中度异常增生、白斑伴上皮重度异常增生及口腔癌中逐渐增加,阳性细胞率分别为(1.05±1.21)%、(6.06 ±4.87)%、(12.49 ±8.41)%和(21.89±10.45)%;caspase-3的表达在3个病例组中逐渐下降,正常对照、白斑伴上皮轻-中度异常增生、白斑伴上皮重度异常增生及口腔鳞状细胞癌组阳性细胞率分别为(12.37 ±5.48)%、(19.51 ±13.15)%、(9.76±7.83)%和(6.08 ±6.91)%;正常对照、白斑伴轻-中度异常增生、白斑伴重度异常增生及口腔鳞状细胞癌组凋亡指数分别为(0.89 ±0.46)%、(1.29 ±0.63)%、(0.65 ±0.40)%和(0.21 ±0.12)%,前3组与口腔鳞状细胞癌组相比差异有统计学意义(P<0.05).结论 生存素在口腔癌的发生过程中起重要作用,随着生存素表达的增加,caspase-3和凋亡指数呈下降趋势;生存索可能通过抑制caspase-3的表达,从而抑制细胞的凋亡,促进口腔癌的发生.     

survivin在口腔鳞状细胞癌中的表达及其与 bcl-2,P53,bax表达之间的关系

目的初步探讨口腔鳞状细胞癌中凋亡抑制蛋白survivin的表达和临床意义,以及survivin的表达与bcl-2,P53,bax表达之间的关系。方法采用免疫组化二步法,DAB染色。27例口腔鳞癌和1例乳腺癌阳性对照病例手术前均未进行放疗、化疗或生物治疗。结果27例口腔鳞癌组织中survivin阳性表达率为88.89%(24/27),11例正常组织中survivin没有表达。在口腔鳞癌不同的临床病理分级survivin的表达有差异(P<0.05),其表达随着临床病理分级的增加而增加。Survivin在有淋巴结转移者阳性表达率(100%)高于无淋巴结转移者(82.35%),但统计学比较无差异(P>0.05)。口腔鳞癌中survivin的表达在不同的年龄组和性别间无差异(P>0.05),而在不同部位的鳞癌中表达有差异(P<0.05)。口腔鳞癌中survivin的表达和bcl-2,突变型P53的表达成正相关,(P<0.05),而与bax的表达成负相关,(P<0.05)。结论①Survivin可作为口腔鳞癌诊断的一项重要指标;②Survivin可作为口腔鳞癌基因治疗和免疫治疗的靶分子;③在口腔鳞癌基因治疗方面可以考虑协同抑制突变型P53和bcl-2的表达,或者提高bax的表达来提高疗效。     

CD1a-positive cells in odontogenic cysts

Langerhans cells (LC) are bone marrow-derived cells that have a CD1a-positive immunophenotype and are an important portion of the cell-mediated immune response. The aim of this study was an immunohistochemical evaluation of CD1a positive cells in different types of oral cysts. Fifty-five cysts were studied: 18 odontogenic keratocysts (OKC), of which five were orthokeratotic and 13 parakeratotic; 19 radicular cysts; and 18 dentigerous cysts. Positive LC was 80% for orthokeratotic OKC, 33% for parakeratotic OKC, approximately 35% for radicular cysts, and approximately 20% for dentigerous cysts. The results show that OKC with well-differentiated epithelial linings presented a greater number of LC than the other cysts. However, when the cyst wall was inflamed there were no differences in LC expression in the different types of cysts. The data confirm that LC distribution seems to be associated with the degree of differentiation of the epithelia.     

Differentiation of odontogenic keratocysts from nonkeratinizing cysts by use of fine-needle aspiration biopsy and cytokeratin-10 staining.

PURPOSE: In this study, the efficacy of fine-needle aspiration biopsy (FNAB) and cytokeratin 10 immunocytochemical staining to differentiate odontogenic keratocysts (OKC) from dentigerous and other nonkeratinizing cysts was evaluated. PATIENTS AND METHODS: This was a prospective study of 18 FNABs of odontogenic cystic lesions performed at the Massachusetts General Hospital between 1995 and 1998. A consistent and standardized technique was used to obtain the cytologic material. Immunocytochemistry was performed on destained smears by using a monoclonal antibody against cytokeratin 10. Identical immunohistochemical methods were applied to the final surgical specimen, and results were compared. RESULTS: Cells of 10 of 18 FNABs showed a markedly positive immunoreaction to anti-cytokeratin 10, supporting a diagnosis of OKC. In all 10 cases, the diagnosis was confirmed by histology. Six of 18 cases showed an absence of staining and were interpreted as anti-cytokeratin 10 negative. In the 2 remaining cases, there were occasional squamous cells on the smear with weak anti-cytokeratin 10 uptake. The overall pattern was negative, and these were interpreted as nonkeratinizing cysts. In all 8 of these cases, the diagnosis of OKC was excluded based on the immunocytochemistry, and the final histologic diagnoses were: dentigerous cyst (n = 4) and radicular cyst (n = 4). CONCLUSIONS: The combination of FNAB with immunocytochemical determination of cytokeratin 10 expression by sampled epithelial cells was 100% accurate in distinguishing an OKC from a nonkeratinizing odontogenic cyst in this series. The technique allows for early diagnosis and rational surgical planning.     

Quantification of PCNA+ cells within odontogenic jaw cyst epithelium

The aim of this study was to investigate the reactivity of the epithelial linings of the three major types of odontogenic cyst with a monoclonal antibody to proliferating cell nuclear antigen (PCNA; clone PC 10). PCNA expression was studied in odontogenic cysts (n=31) and normal oral epithelium (n= 10) using a biotin-streptavidin method on routinely processed paraffin sections. PCNA⁺ cells were counted manually and related to the length of basement membrane (mm) and the epithelial area (mm²) as determined by TV image analysis. The epithelial linings of odontogenic keratocysts (OKC; n= 11) contained the highest number of PCNA⁺ cells, most of which were located in the suprabasal layers. The mean value of PCNA⁺ cells in OKC linings (94.4 ±22.7 cells/mm) was similar to that of oral epithelia (80.8 ± 20.6 cells/mm), but both were significantly higher than that of dentigerous (n = 10. 5.1 ± 3.0 cells mm) and radicular (n = 10, 11.0 ± 4.1 cells /mm) cyst linings (P- 0.005). The epithelial distribution of PCNA⁺ cells differed between groups with the basal/suprabasal PCNA⁺ cell ratio in OKC linings (0.05 ± 0.02) being significantly lower than that of normal oral epithelium (0.5 ± 0.14), dentigerous (l.6 ± 1.23) and radicular (l.9 ± 1.09) cyst linings respectively (P < 0.005). These results demonstrate differences in PCNA⁺ expression between the epithelial linings of the major odontogenic cyst types, indicating differences in proliferative and differentiation processes within these lesions.     

Evaluation of collagen in connective tissue walls of odontogenic cysts –A histochemical study

J Oral Pathol Med (2011) 40 : 257–262 Background: The purpose of this study was to evaluate the nature of collagen in the connective tissue walls of odontogenic cysts, like the odontogenic keratocyst (OKC), dentigerous cyst and radicular cyst using picrosirius red stained sections. Furthermore, it was intended to assess if the capsular connective tissue can affect the nature of overlying epithelium, thus emphasizing the role of epithelial–mesenchymal interactions in biological behaviour of the cysts. Materials and method: The material for the study included 51 formalin‐fixed paraffin‐embedded tissue blocks (15 odontogenic keratocyst, 15 dentigerous cysts, 15 radicular cysts and four normal mucosa and two dental follicular tissue as controls), retrieved from the Department of Oral Pathology and Microbiology, MCODS, Manipal. Tissue blocks were sectioned at 5‐μm thickness, stained with picrosirius red stain and observed with polarization and light microscopy. Results: Few sections of OKC and dentigerous cyst exhibited greenish‐yellow birefringence in sub‐epithelial region, whereas others showed a yellowish‐orange birefringence under polarization microscopy. Most radicular cysts had yellowish‐orange to orange birefringence. Shift in colour in case OKC and dentigerous cyst was attributed to the presence of inflammation in those sections. These regions also exhibited either a change in phenotype or thickness of overlying epithelium. Conclusion: This technique can be used to study the nature of collagen fibres in odontogenic cyst walls. Further studies with an increased sample size and using various epithelial and mesenchymal markers and ssDNA antibodies should be carried out to confirm the effect of epithelial–mesenchymal interactions on the nature of epithelium of odontogenic cysts.     

The immunoprofile of odontogenic keratocyst (keratocystic odontogenic tumor) that includes expression of PTCH, SMO, GLI-1 and bcl-2 is similar to ameloblastoma but different from odontogenic cysts

Background:  The aggressive biological behavior of odontogenic keratocysts (OKCs), unlike that of other odontogenic cysts, has argued for its recent re-classification as a neoplasm, 'keratocystic odontogenic tumor'. Identification of mutations in the PTCH gene in some of the OKCs that were expected to produce truncated proteins, resulting in loss of control of the cell cycle, provided additional support for OKCs having a neoplastic nature.
Methods:  We investigated the immunohistochemical expression of the sonic hedgehog (SHH) signaling pathway-related proteins, PTCH, smoothened (SMO) and GLI-1, and of the SHH–induced bcl-2 oncoprotein in a series of primary OKC (pOKC), recurrent OKC (rOKC) and nevoid basal cell carcinoma syndrome-associated OKCs (NBCCS-OKCs), and compared them to solid ameloblastomas (SAMs), unicystic ameloblastomas (UAMs), 'orthokeratinized' OKCs (oOKCs), dentigerous cysts (DCs) and radicular cysts (RCs).
Results:  All studied lesions expressed the SHH pathway-related proteins in a similar pattern. The expression of bcl-2 in OKCs (pOKCs and NBCCS-OKCs) and SAMs was significantly higher than in oOKCs, DCs and RCs ( P  <   0.001).
Conclusions:  The present results of the immunoprofile of OKCs (that includes the expression of the SHH-related proteins and the SHH-induced bcl-2 oncoprotein) further support the notion of OKC having a neoplastic nature. As OKCs vary considerably in their biologic behavior, it is suggested that the quality and quantity of interactions between the SHH and other cell cycle regulatory pathways are likely to work synergistically to define the individual phenotype and corresponding biological behavior of this lesion.     

细胞凋亡抑制基因bcl-2在牙源性病损中表达的研究

目的　观察凋亡相关基因bcl- 2在成釉细胞瘤 (Ameloblastoma,AB)和牙源性角化囊肿 (OdontogenicKeratocyst,OKC)中的表达 ,探讨其与AB和OKC的生物学意义。方法　应用免疫组化法 (S -P)检测 75例AB(原发 31例 ,复发 37例 ,恶变 7例 ) ,35例OKC及 9例口腔正常粘膜。同时采用原位杂交法检测了 2 0例AB(12例原发 ,8例复发 ) ,12例OKC中的bcl- 2mRNA。结果　 88% (6 6 / 75 )AB ,74.2 % (2 6 / 35 )OKC ,44 .4% (4 / 9)正常口腔粘膜有bcl- 2蛋白表达 ,三组间相比差异有显著性 (P <0 .0 0 1)。同样在原发组AB 77.4% (2 4/ 31) ,复发组AB94.5 % (35 / 37) ,恶性组AB10 0 % (7/ 7)。bcl- 2阳性率及阳性强度随着组织学分级变化而增加 ,组间统计 (P <0 .0 5 )。在 2 0例AB及 12例OKC中bcl- 2mRNA表达趋势与bcl- 2蛋白的表达是相一致的 (P <0 .0 1)。bcl- 2阳性表达与AB不同组织学分型 ,病损发生部位 ,肿瘤生长方式 (单房与多房 )无关 (P >0 .0 5 )。bcl- 2在病变中表达的特征为AB的外周层细胞为强表达 ,星网状层有阳性表达 ,随着细胞增殖分化呈实性片状时bcl- 2表达增强 ,角化退变样细胞 ,颗粒性变细胞表达缺失。OKC中多在基底层细胞中表达。结论　①bcl- 2在AB及OKC的发生、发展及细胞分化与增殖中起着重要作用。     

Immunohistochemical analysis of P53 protein in odontogenic cysts

The p53 is a well-known tumor suppressor gene, the mutations of which are closely related to the decreased differentiation of cells. Findings of studies on immunohistochemical P53 expression in odontogenic cysts are controversial. The present study was carried-out to investigate the immunohistochemical expression of P53 protein in odontogenic cysts. Thirty paraffin blocks of diagnosed odontogenic cysts were processed to determine the immunohistochemical expression of P53 protein. Nine of the 11 odontogenic keratocysts (81.8%) expressed P53, one of three dentigerous cyst cases expressed P53, while none of the 16 radicular cysts expressed P53 protein. The findings of the present work supported the reclassification of OKC as keratocystic odontogenic tumor.     

牙源性角化囊肿与正角化牙源性囊肿CK10、Bcl-2免疫组化表达

目的 :比较牙源性角化囊肿 (odontogenickeratocyst,OKC)与正角化牙源性囊肿 (orthokeratinizedodonto geniccyst,OOC)中CK10及Bcl- 2的表达情况。方法 :OKC及OOC各 10例 ,分别行CK10、Bcl- 2免疫组化染色 ,并利用SPSS10 .0统计软件对免疫组化染色结果进行统计学处理。结果 :CK10在OKC中的阳性表达率为 80 % (8/10 ) ,而在OOC中为 10 0 % (10 / 10 ) (P >0 .0 5 )。OOC上皮中CK10阳性着色于除基底细胞层外的上皮全层 ,而OKC中CK10阳性着色仅见于上皮表层的不全角化层。Bcl- 2在OKC中的阳性表达率为 6 0 % (6 / 10 ) ,而在OOC中为 10 % (1/ 10 ) (P <0 .0 5 )。结论 :OKC与OOC的衬里上皮中免疫组化表达存在显著差别 ,OOC可能为有别于OKC的一种独立病损。     

Odontogenic keratocyst: Review of 256 cases for recurrence and clinicopathologic parameters

Odontogenic keratocyst (OKC) is of particular interest because of its high recurrence rate and aggressive behavior. Two hundred fifty-six cases of OKC were reviewed for the age of the patient at diagnosis, sex of the patient, OKC location, and radiographic findings, and 132 patients with OKC were observed to estimate recurrence, which was analyzed for age, sex, location, and several histopathologic findings. OKCs occurred more frequently in men (58.6%) than in women (41.4%), and they occurred in patients within a wide age range, most commonly in patients in the third decade of life (28.9%), followed by those in the second decade (25.0%); the mean age of patients with OKC was 30.8 years. One hundred ninety-six of the 256 cases (76.5%) occurred in the mandible, and the other 60 cases (23.5%) occurred in the maxilla. The mandibular molar and the premolar areas (51.2%) were the most common sites, and the most frequent clinical manifestations at first admission were swelling, pain, or both (82.4% of total cases). Radiographic impressions included dentigerous cyst (27.3%), OKC (25.4%), primordial cyst (14.8%), ameloblastoma (11.7%), residual cyst (9.8%), and radicular cyst (3.1%). The frequency of recurrence at the follow-up examination was 58.3%. There was no significant difference in the recurrence rate on the basis of the sex of the patient. However, OKCs had a significantly higher recurrence rate in patients in the fifth decade of life than in patients in the other age groups (P = .005).Recurrence rates were significantly dependent on the sites of involvement, and OKCs in the mandibular molar region had significantly higher recurrence rates than those in other sites (P = .001). The histopathologic presence of one or more daughter cysts was significantly related to recurrence (P = .03).