iNOS在口腔扁平苔藓癌变过程中的表达研究

目的 :研究诱导型一氧化氮合酶 (iNOS)在口腔扁平苔藓、口腔鳞状细胞癌发展过程中的表达水平及其作用。方法 :以正常口腔黏膜作对照 ,采用免疫组化SP法检测 2 5例口腔扁平苔藓和 10例口腔鳞状细胞癌组织中的iNOS表达。结果 :正常口腔黏膜iNOS阴性表达 ;扁平苔藓组和鳞癌组iNOS表达较正常黏膜组均非常显著增加 (P <0 .0 1) ;扁平苔藓组与鳞癌组之间以及扁平苔藓组内部按上皮异常增生程度分组之间iNOS的表达均无显著差异 (P >0 .0 5 )。结论 :iNOS在口腔扁平苔藓伴上皮异常增生和鳞癌中高表达 ,在口腔鳞癌的发生发展中起着重要的作用。     

Immunohistopathological study of the oral lichenoid lesions of chronic GVHD

BACKGROUND: Chronic graft-vs.-host disease (cGVHD) is a common and serious complication after bone marrow transplantation (BMT). However, the detailed process of oral lichenoid lesions of cGVHD is still unknown. Therefore, we investigated the immunohistopathological features of cGVHD compared with oral lichen planus (OLP) and healthy controls. METHODS: Nineteen allogenic BMT recipients with a histopathological diagnosis of cGVHD were investigated. We investigated the immunohistopathological features of cGVHD compared with OLP and healthy controls. RESULTS: Immunohistopathological features showed that the infiltrations of CD4-positive T cells of cGVHD and OLP were significantly larger than those of the normal oral mucosa (P < 0.005). A larger number of CD8-positive T cells was infiltrated in cGVHD and OLP compared with the normal oral mucosa (P < 0.001). The difference in the number of CD4- and CD8-positive T cells between cGVHD and OLP was not significant. The infiltrations of Langerhans cells (CD1a) in cGVHD and OLP were significantly larger than those in the normal oral mucosa (P < 0.005). The difference in the number of Langerhans cells between cGVHD and OLP was not significant. CD68-positive macrophages were more frequently seen in cGVHD and OLP than in the normal oral mucosa (P < 0.0001). The difference in the number of CD68-positive macrophages between cGVHD and OLP was not significant. CONCLUSIONS: It is suggested that Langerhans cells and CD8-positive T cell may play a major role in the pathogenesis of the oral lichenoid lesions of cGVHD, and the immune response was inducted in OLP as well as the oral lichenoid lesion of cGVHD in this study.     

VCAM-1 and ICAM-1 are expressed by Langerhans cells, macrophages and endothelial cells in oral lichen planus

Expression of intercellular adhesion molecule-1 (ICAAM-1, CD54) and vascular cell adhesion molecule-1 (VCAM-l, CD106) was examined in oral lichen planus (OLP) and normal oral mucosa (NOM). Immunoperoxidase staining showed ICAM-1 expression by vascular endothelium in all biopsies of OLP and NOM whereas endothelial VCAM-l staining was found in 2/7 NOM and 8/9 OLP. In the lamina propria of NOM occasional cells were ICAM-1 or VCAM-l positive, and virtually no staining of intraepilhelial dendritic cells was seen for either marker. Intraepithelial dendritic cells stained for ICAM-1 in 7/9 and VCAM-1 in 4/9 OLP biopsies. Double immunofluorescence showed dual labelling of Langerhans cells (LC) with CD1a and VCAM-l in a further 5/12 cases of OLP, but there was no such staining in four NOM. This is the first report of LC staining with VCAAM-l. Induction of ICAM-1 and VCAM-l on LC and macrophages in OLP suggests these cells are activated and may contribute to the pathogenesis of OLP by presenting antigen to infiltrating lymphocytes.     

口腔扁平苔藓固有层淋巴细胞中Fas及Fas配体的表达变化

目的 检测Fas及Fas配体(Fas ligand,FasL)在口腔扁平苔藓(oral lichen planus,OLP)固有层淋巴细胞中的蛋白表达,探讨Fas、FasL和活化诱导的细胞死亡(activation-induced cell death,AICD)与OLP发病的关系.方法 采用脱氧核糖核苷酸末端转移酶介导的原位缺口末端标记法检测31例OLP和10例正常口腔黏膜(normal oral mucosa,NOM)中淋巴细胞凋亡情况;分别使用免疫组化法检测组织总淋巴细胞及CD8~+、CD4~+T细胞Fas、FasL的表达.结果 OLP与NOM固有层淋巴细胞凋亡率[分别为(1.9±1.8)%、(11.5±9.0)%]差异有统计学意义(P=0.013).OLP淋巴细胞Fas、FasL表达与NOM组相比明显增强[阳性表达率分别为52%(16/31)、71%(22/31),P值分别为0.005、0.000].OLP中CD8~+与Fas~+细胞双阳性表达率为10%,与NOM组相比无明显升高(P=0.313),而CD8~+与FasL~+细胞双阳性表达率[58%(3/31)]显著升高(P=0.002).CD4~+与Fas~+细胞双阳性表达率为35%(11/31),较NOM组显著升高(P=0.031),其中网纹型的表达明显升高(阳性表达为8/19,P=0.019),但糜烂、萎缩型的表达无显著升高(阳性表达为3/12,P=0.097).CD4~+与FasL~+表达率为16%(5/31),与NOM组相比无明显增强(P=0.182).结论 OLP淋巴细胞凋亡低下;OLP中T细胞亚群Fas、FasL表达不均衡,CD8~+细胞和萎缩、糜烂型OLP中CD4~+细胞可能逃逸AICD,与炎症的持续和进展有关;网纹型OLP中部分CD4~+T细胞可能经历AICD.     

血管内皮生长因子在口腔扁平苔藓及鳞癌组织中的表达

目的探讨血管内皮生长因子在口腔扁平苔藓癌变及发病机制中的作用。方法采用免疫组化法,检测10例正常口腔黏膜、25例口腔扁平苔藓、11例口腔扁平苔藓伴不典型增生及14例口腔鳞癌,上皮组织中血管内皮生长因子的表达水平。结果血管内皮生长因子在24例口腔扁平苔藓标本中阳性表达2例,阴性表达22例,阳性表达率明显低于其它3组（P〈0.05）。扁平苔藓伴不典型增生及口腔鳞癌中血管内皮生长因子的阳性表达率均高于正常黏膜（P〈0.05）。结论血管内皮生长因子的表达异常可能在口腔扁平苔藓的发生发展及癌变的过程中起作用。     

表皮生长因子受体在口腔扁平苔藓组织中的表达

目的探讨口腔扁平苔藓(OLP)癌变及发病机制。方法采用免疫组化法检测10例正常口腔黏膜,16例扁平苔藓,10例扁平苔藓伴不典型增生,14例口腔鳞癌上皮组织中表皮生长因子受体(EGFR)的表达水平。结果OLP伴不典型增生组织中EGFR的表达高于正常黏膜(P<0.05)。口腔鳞癌中EGFR的表达高于正常黏膜及OLP(P<0.01)。结论EGFR的过表达在OLP的发生、发展过程中可能起着重要的作用。     

凋亡蛋白Bcl-2、Bax在白斑、口腔扁平苔藓中的表达

目的:观察凋亡蛋白Bcl-2、Bax在白斑、口腔扁平苔藓上皮细胞中的表达,探讨其在口腔白斑、口腔扁平苔藓癌变过程中的作用机制。方法:采用免疫组化法检测10例正常口腔黏膜上皮、18例口腔扁平苔藓、23例白斑、22例口腔鳞癌上皮组织中凋亡相关蛋白Bcl-2、Bax的表达水平。结果:Bcl-2在白斑、口腔扁平苔藓上皮细胞层无异常表达,但在口腔扁平苔藓淋巴细胞浸润带过度表达。Bcl-2在鳞癌组织中呈高表达,与正常黏膜相比有显著性差异(P<0.05)。Bax在上皮单纯增生、轻度、中度不典型增生和低分化鳞癌及糜烂型口腔扁平苔藓组织中呈过度表达,与正常黏膜相比有显著性差异(P<0.05)。结论:Bax参与了口腔白斑癌变的早期事件,而Bcl-2在不典型增生转化为鳞癌的阶段并未发生作用。口腔扁平苔藓的发病机制可能与Bcl-2抑制淋巴细胞凋亡,使细胞免疫亢进,从而刺激上皮细胞Bax过度表达,诱导角朊细胞凋亡有关。     

缺氧诱导因子-1α在口腔扁平苔藓组织及鳞状细胞癌组织中的表达研究

目的：通过免疫组化方法观察缺氧诱导因子-1a（hypoxia induced factor-1α,HIF-1α）在正常口腔黏膜、口腔扁平苔藓（oral lichen planus,OLP）、OLP伴不典型增生及口腔鳞状细胞癌中的表达,探讨HIF-1α在OLP发生发展以及癌变过程中的作用。方法：收集OLP25例,OLP伴不典型增生11例,口腔鳞状细胞癌14例,正常口腔黏膜10例组织标本,采用免疫组化方法对各组织中HIF-1α表达进行分析。结果：HIF-1α在正常口腔黏膜组织中不表达,在OLP组织、OLP伴不典型组织及口腔鳞状细胞癌组织中均有阳性表达。表达强度由低到高依次为：OLP伴不典型增生组、OLP组、鳞状细胞癌组,各组间均有显著性差异（P〈0.005）。结论：正常口腔黏膜组织,OLP,OLP伴不典型增生以及鳞状细胞癌组织中HIF-1α的表达具有显著性差异,提示HIF-1α参与了口腔扁平苔藓的发生发展以及癌变的过程。     

CD44在口腔扁平苔藓中的表达及意义

目的 探讨口腔扁平苔藓病变中粘附分子CD44H、CD44V3表达的意义。方法 应用免疫组织化学SP法对62例口腔扁平苔藓，其中糜烂型扁平苔藓14例，非糜烂型扁平苔藓48例，21例慢性盘状红斑狼疮，10例正常粘膜CD44H、CD44V3的表达进行观察。结果 扁平苔藓病损区组织的染色以基底细胞、棘层细胞下调明显。结论 CD44参与口腔扁平苔藓的发病过程。     

端粒酶逆转录酶mRNA在OLP和OSCC中的表达

目的：探讨端粒酶逆转录酶（TERT）mRNA在口腔扁平苔藓（OLP）和口腔鳞状细胞癌（OSCC）中的表达以及在OLP癌变过程中的作用。方法：应用mRNA原位杂交法对正常口腔黏膜12例,非糜烂型OLP20例,糜烂型OLP20例及OSCC20例,进行hTERTmRNA的检测。结果：正常口腔黏膜、非糜烂型OLP、糜烂型OLP及OSCC中hTERTmRNA阳性表达率分别为8.33％（1／12）、15％（3／20）、45％（9／20）、80％（16／20）。其中,除正常口腔黏膜与非糜烂型OLP中hTERT mRNA阳性率无显著性差异外,其余各组间均有显著性差异（P〈0.05）。而且,正常口腔黏膜与非糜烂型OLP的hTERT mRNA染色强度明显低于糜烂型OLP及OSCC（P〈0.05）。结论：hTERT mRNA可能在糜烂型OLP的癌变过程起着一定的作用。     

诱导型一氧化氮合酶表达在口腔癌前病变口腔鳞癌发展过程中的作用研究

目的　探讨诱导型一氧化氮合酶 (iNOS)在口腔癌前病变、口腔鳞癌发展过程中的表达情况及其作用。方法　采用免疫组化SABC法检测 10例正常口腔黏膜、8例上皮单纯增生、2 0例上皮异常增生和 3 2例鳞状细胞癌组织中iNOS的表达。结果　正常口腔黏膜iNOS阴性表达 ;上皮异常增生组和鳞癌组中iNOS的表达较上皮单纯增生组均显著增加 (P <0 .0 5 ) ;随着上皮异常增生程度的加重 ,iNOS的标记指数逐级显著上升 (P <0 .0 5 ) ;上皮异常增生组与鳞癌组之间以及鳞癌的各病理分级之间iNOS的表达均无显著性差异 (P >0 .0 5 )。结论　iNOS的表达可能参与了口腔鳞癌的衍进过程     

口腔癌前病变细胞凋亡状况及Bcl-2、Bax表达的研究

目的 :通过对口腔白斑、扁平苔藓病变发展及癌变过程中细胞凋亡状况和凋亡相关蛋白表达水平的研究 ,探讨口腔白斑、扁平苔藓的癌变机理及病变机制。方法 :采用原位末端转移酶标记法及免疫组化法 ,观察分析 10例正常口腔黏膜上皮 ,18例扁平苔藓 ,2 3例白斑 ,2 2例口腔鳞癌上皮组织中细胞凋亡状况及凋亡相关蛋白Bcl- 2、Bax的表达水平。结果 :上皮 (轻 ,中 ,重 )不典型增生 ,鳞癌及糜烂型扁平苔藓的凋亡指数均高于正常 (P <0 .0 1)。Bcl- 2在白斑、扁平苔藓上皮细胞层无异常表达 ,但在扁平苔藓固有层淋巴细胞浸润处过度表达 ,在鳞癌组织中显高表达 (P <0 .0 1)。Bax在上皮单纯增生、轻、中度不典型增生和鳞癌及糜烂型扁平苔藓组织中显过度表达 (P <0 .0 5 )。结论 :白斑、扁平苔藓的病变发展及癌变过程中 ,上皮细胞凋亡状况发生了改变 ,Bax参与了白斑癌变的早期事件。扁平苔藓的发病机制与Bcl- 2、Bax在特定部位的过度表达有关     

Expression of keratins 8, 18, and 19 in epithelia of atrophic oral lichen planus

Keratins form intermediate filaments of the cytoskeleton in keratinocytes and have roles in cell structure, signaling, intracellular transport, and cell death. Oral lichen planus (OLP) is an oral inflammatory disease with derangements in basal keratinocytes and disruption of the basal membrane. Here, we focused on epithelial expression of keratins 8, 18, and 19 because these proteins are known to modulate cell death. Biopsies were taken from buccal oral mucosa of persons with normal oral mucosa (n = 10) or atrophic OLP (n = 10). Cultured normal oral keratinocytes (n = 4) showed expression of mRNA and protein for keratins 8, 18, and 19. Immunohistochemistry showed consistent staining for keratins 8 and 18 in basal keratinocytes of normal oral mucosa. In OLP, staining for keratin (K)8 was mostly negative and staining for K18 was weak. Keratin 19 was expressed irregularly in most biopsies of normal oral mucosa and not at all in OLP. Several mononuclear leukocytes in the cellular infiltrate showed membrane staining for K8 and K18. Positive staining for K16 confirmed partial collapse of the basal cell layer in OLP. The basal cell niche in OLP therefore appeared to be partly populated with keratinocytes demonstrating a higher degree of differentiation (K8⁻ K18⁻ K19⁻ K16⁺); consequently, such areas may be more susceptible to the action of cell death factors released from the cell infiltrate as a result of lacking the protective, normal keratin present in the basal epithelial cell layer of normal oral mucosa.     

Expression of inducible nitric oxide synthase and p53 in oral epithelial dysplasia

AIM: Nitric oxide (NO) has been studied in a variety of human cancers and is implicated in both tumor promotion and inhibition. Downregulation of the enzyme iNOS by wild-type p53 (but not mutant) protein has been shown to occur in normal cells and some tumors, but the relationship has not been reported in oral epithelial dysplasia. METHODS AND RESULTS: An immunohistochemical study was conducted with antibodies to iNOS and p53 (clone DO-7) in 36 cases of oral dysplasia of varying severity. Statistical analysis showed a significant correlation between iNOS staining and grade of dysplasia (P <.001) and between p53 and iNOS staining (P <.001). CONCLUSIONS: This preliminary study has shown that iNOS expression correlates with severity of dysplasia, and it is also increased in those cases showing positive staining for p53. Further research is required to fully establish the relationship between iNOS and p53 in both dysplasia and oral squamous cell carcinoma.     

Does expression of inducible nitric oxide synthase correlate with severity of oral epithelial dysplasia?

The small molecule nitric oxide (NO) has generated an exponential amount of research since its discovery as a biological messenger in 1987. It has a vast number of actions, many of which are poorly understood. It has been studied in a variety of human cancers and has been implicated both in tumour promotion and inhibition. Although NO is produced by three distinct isoforms of the enzyme nitric oxide synthase (NOS), most cancer research is directed towards the calcium-independent form, iNOS which following induction, produces much higher quantities of NO than the other two. In this study the expression of iNOS is assessed by immunohistochemistry in 26 cases of oral epithelial dysplasia ranging in severity from mild to severe. iNOS staining was found in all 26 cases of dysplasia with the degree of staining correlating to the severity of dysplasia (p < 0.001). There was no iNOS staining seen in adjacent normal epithelium. The possible role of iNOS in the complex transformation from dysplasia to invasive oral cancer and the clinical applications are discussed.     

Heat shock protein expression in oral lichen planus

To assess the potential role of heat shock protein (HSP) in the pathogenesis of oral lichen planus (OLP), sections of OLP, normal oral mucosa, non-specific oral ulceration (NSOU) and dysplastic OLP were assessed for HSP expression using avidin-biotin complex immunohistochemistry with an anti-HSP 70 polyclonal antibody. There were statistically significant differences in both the vertical and horizontal staining distribution when other groups were compared with the OLP group (p<0.01). Using microdensitometry, the mean staining intensity in OLP, dysplastic OLP and NSOU was elevated in comparison with normal oral mucosa (p<0.001). In a standard tritiated thymidine uptake assay, lymphocytes extracted from nine OLP lesions demonstrated significant proliferation when stimulated with purified protein derivative (PPD), of which HSP is a major constituent, with stimulation indices ranging from 2 to 132. These results are consistent with the hypothesis that, in OLP patients, diverse exogenous agenst may cause upregulated expression of HSP by oral mucosal keratinocytes. A reaction of cytotoxic T lymphocytes to these activated keratinocytes may then result in the tissue destruction which is characteristic of OLP lesions.     

口腔扁平苔藓中细胞角蛋白19的表达及意义

目的:探讨细胞角蛋白19(keratin 19,CK19)在口腔扁平苔藓(OLP)中的表达以及其病理意义.方法:采用免疫组化法检测17例OLP和14例正常颊黏膜中CK19的表达,采用RT-PCR方法分别在OLP患者和正常人颊黏膜中比较CK19 mRNA的表达.结果:正常颊黏膜中CK19均在基底细胞胞浆内呈强阳性表达,只有5例(29.4%)OLP基底细胞呈弱阳性表达,其余为阴性表达;RT-PCR结果显示在OLP组织中CK19 mRNA表达显著高于正常黏膜(P＜0.05).结论:CK19的表达下调与颊部OLP黏膜上皮更新能力下降密切相关.     

Oral lichen planus has a high rate of TP53 mutations. A study of oral mucosa in icelanD

Oral squamous cell carcinoma (OSCC) is a world-wide health problem. In addition to external exposure (smoking and alcohol), certain oral lesions may increase the risk of oral cancer (e.g. leukoplakia, erythroplakia, and oral lichen planus). TP53 has been implicated in OSCC, but there are limited studies of mutations in premalignant oral lesions. In this study, 55 samples from OSCC, 47 from hyperkeratotic (HK) oral mucosa, clinically diagnosed as white patches, 48 samples from oral lichen planus (OLP), and 12 biopsies from normal oral mucosa were studied immunohistochemically for expression of TP53 protein. From all the carcinoma samples and selected non-malignant samples showing moderate or strong TP53 protein expression, malignant cells or TP53-positive nuclei were microdissected and screened for mutations in exons 5-8 by constant denaturation gel electrophoresis. Moderate to strong TP53 protein staining was seen in 56% of OSCC, 32% of OLP but only in 13% of HK. All OLP samples showed a characteristic pattern of positive nuclei confined to the basal layer, whereas TP53 staining was seen in suprabasal nuclei in HK. Mutation rate was 11 out of 52 for OSCC, three out of 20 tested for HK and, remarkably, nine out 27 tested for OLP. There was no correlation between TP53 protein staining and TP53 mutations. No associations were found with anatomical sites or disease progression. The unexpectedly high mutation rate of OLP might explain the premalignant potential of this lesion.     

CD133在口腔扁平苔藓和口腔鳞状细胞癌中的表达及其临床意义

目的探究肿瘤干细胞标记物CD133在口腔正常黏膜、口腔扁平苔藓（OLP）及口腔鳞状细胞癌（OSCC）中的表达情况及临床意义,评估其作为OLP恶性转化的早期诊断指标、OSCC治疗干预靶标的临床价值,为进一步研究口腔黏膜癌变机制提供基础。 方法回顾性分析10例正常口腔黏膜、60例OLP、60例OSCC患者的临床资料,运用免疫组织化学技术检测各组病理组织中CD133表达情况,采用Mann-Whiney秩和检验比较各组间CD133表达差异,卡方检验统计分析CD133与各临床因素的关系。采用免疫组织化学技术和免疫印迹技术检测人口腔癌前病变细胞株（DOK）和人OSCC细胞株（CAL-27）中CD133的表达情况,t检验比较CD133在DOK与CAL-27细胞株中含量差异。 结果口腔正常黏膜、OLP、OSCC三组中,CD133的阳性率为0（0/10）、31.67%（19/60）、63.33%（38/60）,表达逐渐增强。CD133在口腔正常黏膜与OLP表达强度差异有统计学意义（Z=-2.046,P= 0.041）。CD133在OLP与OSCC表达强度差异具有统计学意义（Z=-3.777,P<0.001）。CD133在DOK中弱阳性表达,在CAL-27中阳性表达,DOK与CAL-27中CD133含量差异有统计学意义（t=-5.029,P= 0.001）CD133与OSCC的临床分期和淋巴结转移有关。 结论CD133作为评估OLP恶性转化潜能的指标及OSCC早期治疗干预靶标可能具有重要临床价值。     

Overexpression of cdk4 and p16 in oral lichen planus supports the concept of premalignancy

J Oral Pathol Med (2011) 40 : 294–299 Background: Cell cycle arrest and increased cell proliferation have been demonstrated in oral lichen planus (OLP). This study evaluated the expression of cdk4, cdk6 and p16, important proteins in the G1 phase, in OLP and compared the expression of these proteins of OLP with those of normal mucosa. Methods: Expression of cdk4, cdk6 and p16 were investigated in 23 OLP and 10 normal mucosae using immunohistochemistry technique. Positive cells were counted and presented as a percentage of positive cells. Results: Expression of cdk4, cdk6 and p16 was observed in 3/10 (30%), 1/10 (10%) and none of normal mucosa, respectively. Expression of cdk4, cdk6 and p16 was detected in 18/23 (78.3%), 8/23 (34.8%) and 15/23 (65.2%), of OLP, respectively. The numbers of cdk4 and p16 positive cases of OLP were significantly higher than normal mucosa. In normal mucosa, the averages of the percentage of positive cells for cdk4 and cdk6 staining were 1.48 and 0.18, respectively. In OLP, the averages of the percentage of positive cells for cdk4, cdk6 and p16 staining were 2.73, 1.06 and 2.24, respectively. The percentage of cdk4‐positive cells of OLP was significantly higher than those of normal mucosa group. Conclusion: Oral lichen planus demonstrated overexpression of cdk4 and p16, but not cdk6, suggesting that epithelial cells in OLP are in the hyperproliferative state and in cell arrest. Altered expression of cdk4 and p16 provides evidence of the malignant potential in OLP.