PCR-SSCP和DNA测序检测牙源性角化囊肿中PTCH基因的突变

目的 检测牙源性角化囊肿（OKC）中PTCH基因突变的特点。方法 采用PCR-SSCP筛查与DNA直接测序的方法对12例OKC进行PTCH基因突变的检测, 其中2例为痣样基底细胞癌综合征（NBCCS）相关OKC,10例为散发OKC。 结果 在4例OKC中发现了4处突变,其中2处生殖细胞突变发生在2例NBCCS相关OKC,2处体细胞突变发生在2例散发OKC。另外,还在10例OKC中检测到了8处PTCH基因多态性。结论 NBCCS相关OKC和散发OKC均可发生PTCH基因突变,但突变水平不同,PTCH基因的突变在二者的发病中可能均起重要作用。     

牙源性角化囊肿中PTCH基因的突变检测

目的检测牙源性角化囊肿（OKC）中PTCH基因突变的发生频率、类型及分布特点，分析散发OKC与伴发痣样基底细胞癌综合征（NBCCS）OKC之间的分子病理联系。方法收集8例OKC新鲜病变组织（4例散发，4例伴发NBCCS），提取DNA，采用PCR直接测序法检测OKC病变组织中的PTCH基因突变。结果分别于4例NBCCS—OKC和2例散发OKC中检测到6处PTCH基因突变，2例为错义突变，引起1个氨基酸的改变；其余4例突变分别为1～7个碱基插入或缺失，其中3例引起读码框的改变（移码突变），并导致蛋白质的提前截断，1例导致了2个氨基酸的插入。结论PTCH基因突变不仅常见于NBCCS—OKC，部分散发OKC病变也可以发生该基因的异常。     

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。     

正角化牙源性囊肿的临床病理及免疫组化研究

目的：探讨一组正角化牙源性囊肿（orthokeratinized odontogenic cyst,OOC)的临床病理及免疫组化特点。方法：以OOC的名称报告20例，观察其组织学和免疫组化特点，并与牙源性角化囊肿（odontogenic keratocyst,OKC)病变进行比较。结果：本组病例约占同期所有OKC的9．9％（20／202），其中男性14例，女性6例，就诊平均年龄39．1％；随访资料显示：15例患者行囊肿刮治后无复发；组织学和免疫组化比较发现：OOC和OKC之间差异有显著性，OOC上皮不表现OKC上皮的形态分化特点，细胞增殖活性较低。结论：OOC可能代表一组有别于牙源性角化囊肿的颌骨病损。     

The Odontogenic Jaw Cysts： Analysis of the Clinical Parameters of 669 Cases

目的分析、比较三型牙源性颌骨囊肿的临床特点.方法收集20年间牙源性角化囊肿(odontogenic keratocyst,OKC)、根端囊肿(radicular cyst,RC)及含牙囊肿(dentigerous cyst,DC)的临床资料,对其性别构成、年龄分布、发病部位及临床表现等进行比较研究.结果1)三型颌骨囊肿的男女之比分别为OKC 1.6∶1,RC 1.4∶1,DC 4.1∶1(χ2检验,P＜0.005).2)除DC未见于70岁以上年龄段外,几乎各年龄段均见三型颌骨囊肿的发生,三型囊肿组间及组内的年龄分布均有显著性差异(χ2检验,P＜0.005).OKC及RC20～29岁年龄段患病人数最多,分别占各年龄段患病人数的27%及20%;DC10～19岁年龄段患病人数最多,占各年龄段患病人数的29%.3)颌骨的任一部位均见三型颌骨囊肿的发生,但发生频率不同,三型颌骨囊肿组间及组内发病部位的分布有显著性差异(χ2检验,P＜0.005).4)OKC有137例合并感染,感染率39%;RC48例合并感染,感染率24%;DC18例合并感染,感染率16%,三型间有显著性差异(χ2检验,P＜0.005).结论1)男性较女性更易发生牙源性颌骨囊肿.2)不同的年龄段,对OKC、RC及DC的易感性不同.OKC及RC发生的高峰期均为20～29岁年龄段;DC发生的高峰期为 10～19岁年龄段.3)不同的颌骨部位,对OKC、RC及DC的易感性不同.OKC好发于下颌磨牙区,其次为下颌双尖牙区;RC及DC则好发于上颌前牙区.4)感染症状的出现,对OKC、RC及DC彼此间的鉴别诊断具一定临床意义.     

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。     

端粒酶逆转录酶基因在牙源性病损中的表达

目的 研究成釉细胞瘤（ameloblastoma,AB)和牙源性角化囊肿（odontogenic keratocyst,OKC)中hTERT mRNA的表达，探讨其在AB、OKC中的发生、发展及其生物学意义。方法 采用mRNA原位杂交法对口腔颌骨内54例AB、16例OKC及7例口腔正常粘膜，进行了hTERT mRNA的检测。结果 94.4%(51/54)AB、87.5%(14/16)OKC及1／7正常口腔粘膜有hTERT mRNA表达，3组相比差异有显著性（P＜0.001)。hTERT与AB不同组织学类型、病损发生部位、单房与多房无关（P＞0.05)，在AB外周细胞及星网状多边形细胞中有表达，在角化退变样细胞及颗粒样细胞中缺乏表达。结论 ①hTERT活化在AB及OKC的发生、发展中起重要作用；②hTERT活性阳性率与病变中的细胞分化，临床生物学行为有关，随着组织学分级增高hTERT mRNA阳性率及信号强度均增高。     

成釉细胞瘤及牙源性角化囊肿中ICAM-1和VCAM-1的表达

目的探讨细胞间黏附分子-1(ICAM-1)和血管细胞黏附分子-1(VCAM-1)在成釉细胞瘤(AB)及牙源性角化囊肿(OKC)中的表达及其与AB、OKC病理学特征的关系。方法对38例AB、10例OKC、7例正常口腔黏膜(NOM)组织进行免疫组织化学SP法检测,结合病例病理特征进行分析。结果ICAM-1和VCAM-1在AB、OKC和NOM3组表达组间比较,具有显著统计学差异(P<0.05)。ICAM-1在AB中的阳性率达65.2%,显著高于NOM(14.3%),OKC(60.0%)与NOM未见显著统计学差异。VCAM-1在AB中的阳性血管数也显著高于OKC和NOM。ICAM-1和VCAM-1表达与AB的组织病理分型、年龄、性别和发生部位无明显相关性(P>0.05)。结论细胞黏附分子ICAM-1和VCAM-1与AB及OKC的发生、发展及细胞分化与增殖有关。     

细胞角蛋白18及其基因在牙源性角化囊肿衬里上皮中表达的意义

目的检测细胞角蛋白18（CK18）及其基因在牙源性角化囊肿（OKC）衬里上皮中的表达。方法选取32例OKC的衬里上皮组织,分别进行CK18、CK8和CK19单克隆抗体的免疫组织化学染色。对其中12例使用RT- PCR法检测CK18 mRNA,观察其在衬里上皮中的表达;同时使用CK18基因探针进行原位杂交,检测CK18 mRNA在衬里上皮细胞层的定位表达。结果在免疫组织化学染色中, 17例CK18蛋白在OKC衬里上皮的表层细胞层表达为弱阳性;27例CK18蛋白在棘细胞层上层染色为阳性;14例CK18蛋白在棘细胞层染色为阳性;所有标本基底细胞层染色呈阴性。RT- PCR法检测见4例CK18 mRNA表达为强阳性,8例表达为弱阳性。原位杂交法检测见8例CK18 mRNA在棘细胞层和棘细胞层上层呈阳性,4例在上皮基底细胞层和角化层呈阳性。CK8蛋白在所有32例OKC衬里上皮基底细胞层均有表达。CK19蛋白在23例OKC衬里上皮表层均有表达。结论CK18在OKC衬里上皮的表达由基底细胞层向棘细胞层迁移,CK18蛋白免疫组织化学染色阳性表达与CK18 mRNA原位杂交法阳性表达不同,提示CK18可能与衬里上皮的增殖活性有关,OKC衬里上皮中可能存在CK18蛋白和CK18 mRNA表达的调控因子。     

PTCH germline mutations in Chinese nevoid basal cell carcinoma syndrome patients

OBJECTIVES: PTCH, the human homologue of the Drosophila segment polarity gene, patched, has been identified as the gene responsible for nevoid basal cell carcinoma syndrome. The aim of this study was to investigate PTCH gene mutation in Chinese patients with nevoid basal cell carcinoma syndrome. MATERIALS AND METHODS: DNA was isolated from both odontogenic keratocyst tissue and peripheral blood of five patients with syndrome and one patient with only multiple odontogenic keratocysts, and mutational analysis of the PTCH gene performed by direct sequencing after amplification of all 23 exons by polymerase chain reaction (PCR). RESULTS: A previously reported germline mutation (c.2619C>A) was identified in two familial cases involving the mother and the daughter, with the mother also carrying a novel somatic mutation (c.361_362insGAGC). Three novel germline PTCH mutations (c.1338_1339insGCG, c.331delG and c.1939A>T) were detected in three unrelated patients with syndrome. The patient with multiple odontogenic keratocysts who failed to fulfill the diagnostic criteria of the syndrome also carried a novel germline mutation (c.317T>G). CONCLUSION: The frequent germline PTCH mutations detected in our series provide further evidence for the crucial role of PTCH in the pathogenesis of nevoid basal cell carcinoma syndrome in Chinese.     

PTCH1 and SMO gene alterations in keratocystic odontogenic tumors

Keratocystic odontogenic tumors (KCOTs, previously known as odontogenic keratocysts) are aggressive jaw lesions that may occur in isolation or in association with nevoid basal cell carcinoma syndrome (NBCCS). Mutations in the PTCH1 (PTCH) gene are responsible for NBCCS and are related in tumors associated with this syndrome. Mutations in the SMO gene have been identified in basal cell carcinoma and in medulloblastoma, both of which are features of NBCCS. To clarify the role of PTCH1 and SMO in KCOTs, we undertook mutational analysis of PTCH1 and SMO in 20 sporadic and 10 NBCCS-associated KCOTs, and for SMO, 20 additional cases of KCOTs with known PTCH1 status were also included. Eleven novel (1 of which occurred twice) and 5 known PTCH1 mutations were identified. However, no pathogenic mutation was detected in SMO. Our findings suggest that mutations are rare in SMO, but frequent in PTCH1 in sporadic and NBCCS-associated KCOTs. Abbreviations: NBCCS, nevoid basal cell carcinoma syndrome; KCOTs, keratocystic odontogenic tumors; BCCs, basal cell carcinomas.     

PTCH gene mutations in odontogenic keratocysts

An odontogenic keratocyst (OKC) is a benign cystic lesion of the jaws that occurs sporadically or in association with nevoid basal cell carcinoma syndrome (NBCCS). Recently, the gene for NBCCS was cloned and shown to be the human homologue of the Drosophila segment polarity gene Patched (PTCH), a tumor suppressor gene. The PTCH gene encodes a transmembrane protein that acts in opposition to the Hedgehog signaling protein, controlling cell fates, patterning, and growth in numerous tissues, including tooth. We investigated three cases of sporadic odontogenic keratocysts and three other cases associated with NBCCS, looking for mutations of the PTCH gene. Non-radioactive single-strand conformational polymorphism and direct sequencing of PCR products revealed a deletion of 5 base pairs (bp) in exon 3 (518delAAGCG) in one sporadic cyst as well as mutations in two cysts associated with NBCCS, a nonsense (C2760A) and a missense (G3499A) alteration. This report is the first to describe a somatic mutation of PTCH in sporadic odontogenic keratocysts as well as two novel mutations in cysts associated with NBCCS, indicating a similar pathogenesis in a subset of sporadic keratocysts.     

遗传性牙本质发育不全II型家系DSPP基因突变的检测与分析

目的: 对2个汉族遗传性牙本质发育不全II型家系中的DSPP基因突变进行检测。方法: 对2个遗传性牙本质发育不全II型( DGI-II) 家系的成员进行全身基本情况及口腔专科检查,拍摄口内照,全景片,以及牙片,采集外周静脉血并抽提基因组DNA,应用PCR及DNA 测序技术,结合序列分析方法,对2个家系共15名家系成员(其中患者10名)的外周静脉血基因组DNA牙本质涎磷蛋白(DSPP)基因1~5号外显子及其邻近序列进行序列分析。结果: 家系A中的患者在DSPP的第2外显子发生错义突变c.50C>T(p.P17L);家系B的患者在DSPP第4外显子发生错义核苷酸改变c.506A>G(p.D169G)。结论: DSPP基因突变可能是导致两个DGI-II家系发病的分子基础。[关键词] 牙本质发育不全II型( DGI-II) 牙本质涎磷蛋白(DSPP) 基因突变     

A comparative immunohistochemical analysis of cortactin in orthokeratinized odontogenic cyst (OOC), sporadic odontogenic keratocyst (OKC), and syndromic OKC

ObjectivesTo evaluate and compare the immunohistochemical expression of cortactin in the epithelial lining of orthokeratinized odontogenic cyst (OOC), sporadic odontogenic keratocyst (OKC), and syndromic OKC.MethodsFormalin-fixed paraffin-embedded tissue blocks of histopathologically diagnosed cases of OOC, OKC, syndromic OKC, normal buccal mucosa (NBM), and oral squamous cell carcinoma (OSCC) were examined for immunohistochemical expression of cortactin. Clear brown cytoplasmic and membranous staining was considered positive.ResultsA statistically significant difference was observed between OOC and syndromic OKC (p < 0.001), as well as between sporadic OKC and syndromic OKC (p < 0.001). Although not statistically significant, the expression of cortactin was slightly higher in the basal layer of NBM (mean = 0.47), OOC (mean = 0.27), sporadic OKC (mean = 0.47) syndromic OKC (mean = 1.53), and OSCC (mean = 0.67) than in the parabasal layers of NBM (mean = 0.27), OOC (mean = 0.20), sporadic OKC (mean = 0.47), syndromic OKC (mean = 1.27), and OSCC (mean = 0.60).ConclusionThe expression of cortactin in the basal layer may suggest the formation of invadopodia in the basal layer where the invasion mechanism occurs. This finding is further supported by the higher localization of cortactin in areas of epithelial budding and daughter cysts in syndromic OKC, thereby reaffirming its possible association with recurrence.     

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.     

Keratocystic odontogenic tumour: reclassification of the odontogenic keratocyst from cyst to tumour

The purpose of this paper is to review the features and behaviour of the odontogenic keratocyst (OKC), now officially known as the keratocystic odontogenic tumour (KCOT); to analyze a series of histologically confirmed KCOT cases; and to review and discuss the redesignation of KCOT and the implications for treatment. Redesignation of the OKC as the KCOT by the World Health Organization (WHO) is based on the well-known aggressive behaviour of this lesion, its histology and new information regarding its genetics. Abnormal function of PTCH, a tumour suppressor gene, is noted to be involved in both nevoid basal cell carcinoma syndrome and sporadic KCOTs. Normally, PTCH forms a receptor complex with the oncogene SMO for the SHH ligand. PTCH binding to SMO inhibits growth-signal transduction. SHH binding to PTCH releases inhibition of the signal transduction pathway. If normal functioning of PTCH is lost, the proliferation-stimulating effects of SMO are permitted to predominate. A review of the literature was conducted and results were tabulated to determine whether treatment modality is related to recurrence rate. More aggressive treatment - resection or enucleation supplemented with Carnoy"s solution with or without peripheral ostectomy - results in a lower recurrence rate than enucleation alone or marsupialization. Notably, the recurrence rate after marsupialization followed by enucleation is not significantly higher than that following the so-called aggressive modalities. Our case series consists of 21 patients treated for KCOTs. Results were organized to demonstrate recurrence as it relates to size of lesion and time since treatment and incidence as it relates to patient age and location in the jaws. In our series, the average KCOT surface area measured radiographically was 14 cm². Most lesions were within the 0-15 cm² range and lesions in this range resulted in the greatest number and proportion of recurrences. The recurrence rate of 29% in our case series was consistent with previously established data; all recurrences occurred within 2 years post-intervention. The incidence of primary lesions was highest in the age group 70-79 years; most lesions occurred in the posterior mandible. WHO"s reclassification of the OKC as the KCOT based on behaviour, histology and genetics underscores the aggressive nature of the lesion and should motivate clinicians to manage the disease in a correspondingly aggressive manner. The most effective interventions for the KCOT are either enucleation with Carnoy"s solution, or marsupialization with later cystectomy. Future treatment may involve molecular-based modalities, which may reduce or eliminate the need for aggressive surgical management.