基质金属蛋白酶-1在人涎腺腺样囊性癌细胞中的表达

目的:测定基质金属蛋白酶-1(MMP-1)在人涎腺腺样囊性癌不同转移力细胞系中的表达。方法:以人涎腺腺样囊性癌ACC-2细胞系及其高转移株ACC-M作为研究肿瘤转移分子机制的模型,采用免疫组化法和蛋白印迹(Western blot)法检测MMP-1的蛋白表达水平。结果:MMP-1在ACC-M细胞中的表达水平高于ACC-2细胞。结论:MMP-1在人涎腺腺样囊性癌的侵袭转移过程中可能发挥作用。     

NCAM基因对人涎腺腺样囊性癌细胞系SACC-83生长抑制作用的研究

目的 探讨NCAM基因对人涎腺腺样囊性癌细胞株SACC-83体外生长的抑制作用.方法 应用电穿孔转染方法将NCAMcDNA转入人涎腺腺样囊性癌细胞系SACC-83中,经G418筛选2～3周,用兔抗人NCAM抗体进行免疫组化染色鉴定,然后绘制细胞生长曲线,进行软琼脂培养.结果 应用电穿孔方法可成功将NCAM基因转染入人涎腺腺样囊性癌细胞株SACC-83中,免疫组化染色鉴定显示转染NCAM的SACC-83细胞有NCAM蛋白的表达.与对照组相比,经NCAM基因转染后的SACC-83细胞增殖明显受到抑制.转染了NCAM基因的SACC-83细胞克隆形成率明显降低.结论 NCAM基因对人涎腺腺样囊性癌细胞系SACC-83的生长有一定的抑制作用.     

EphA2在涎腺肿瘤中的表达及其临床意义

目的检测EphA2在涎腺肿瘤中的表达，并分析其临床病理意义。方法免疫组化S-P法检测10例正常涎腺组织、30例多形性腺瘤、30例黏液表皮样癌（MEC）、30例腺样囊性癌（ACC）中EphA2的表达。分析EphA2表达与患者的年龄、性别、肿瘤部位、大小、TNM分期以及复发、淋巴结转移等临床病理因素的关系。比较EphA2在正常涎腺组织及几种常见的涎腺良、恶性肿瘤中的表达差异。结果EphA2在正常涎腺组织中主要表达于腺管上皮，而腺泡细胞几乎不表达。EphA2在正常涎腺组织、多形性腺瘤、涎腺癌（包括黏液表皮样癌和腺样囊性癌2组）中的表达依次增高，存在显著性差异（P〈0．05）。EphA2的表达与涎腺癌瘤体大小、TNM分期、复发和淋巴结转移有关（P〈0．05）。结论EphA2可能影响涎腺肿瘤发生、发展及其预后。     

Dermo-1在涎腺腺样囊性癌中的表达及其临床意义

目的探讨转录因子Dermo-1在涎腺腺样囊性癌中的表达水平及其与各临床病理因素的关系。方法采用免疫组化方法检测40例涎腺腺样囊性癌组织中Dermo-1蛋白的表达情况,分析Dermo-1表达与腺样囊性癌临床病理因素的关系,以20例正常涎腺组织作对照。结果 Dermo-1在涎腺腺样囊性癌组织中的表达水平明显高于正常涎腺组织（P〈0.05）。Dermo-1表达与腺样囊性癌的病理分型、嗜神经侵袭、术后复发及远位转移有关（P〈0.05）。结论 Dermo-1蛋白表达可能与涎腺腺样囊性癌的分化调节、嗜神经侵袭及远位转移有关系,检测Dermo-1的表达水平对判断腺样囊性癌患者的预后有一定的参考价值。     

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目的探讨肌动蛋白聚合蛋白-1(Fascin-1)在人涎腺腺样囊性癌(SACC)中的表达及临床意义。方法选择2007年11月至2009年12月哈尔滨医科大学附属口腔医院口腔颌面外科手术切除涎腺组织的患者63例,其中SACC患者46例,涎腺正常患者17例。应用免疫组化法检测SACC和正常涎腺组织中Fascin-1的表达水平,比较并分析其与患者其他因素的相关性。结果 Fascin-1在SACC组织中的表达率明显高于正常涎腺组织(P<0.01)。Fascin-1在SACC组织中的表达,与患者的年龄、性别、部位、肿瘤大小和病理类型无关(P>0.05)。临床分期Ⅲ～Ⅳ期的癌组织Fascin-1表达阳性率明显高于Ⅰ～Ⅱ期(P<0.05),有区域淋巴结转移患者Fascin-1表达阳性率明显高于无区域淋巴结转移者(P<0.05)。结论 Fascin-1表达与SACC患者的临床分期和淋巴结转移相关。     

细胞外基质与涎腺腺样囊性癌的远处转移

目的 研究涎腺腺样囊性癌与细胞外基质的关系。方法 通过肿瘤手术标本的免疫组化染色和超微结构观察、细胞系体外粘附实验及人工重组基底膜侵袭实验，并采用整合素受体抑制剂精氨酸-天冬氨酸进行体外及荷瘤动物体内实验，观察其预防及治疗肺转移的效果。结果 肿瘤团索周围的基膜样物质呈多层、溶解或断裂现象。出现腺样囊性癌远处转移患者的肿瘤标本组织蛋白酶D免疫组化阳性反应率(75％)明显高于无转移患者(43．8％)。肺高转移涎腺腺样囊性癌细胞株对人工重组基膜的侵袭细胞数明显高于其相应的非高转移细胞系。精氨酸-天冬氨酸在体外能抑制肺高转移涎腺腺样囊性癌细胞对人工重组基膜的侵袭，在体内能显著延长涎腺腺样囊性癌实验性肺转移动物的生存期。结论 涎腺腺样囊性癌的远处转移与细胞外基质有密切关系。     

涎腺腺样囊性癌组织中层粘连蛋白及血管内皮生长因子表达的研究

目的 :探讨涎腺腺样囊性癌层粘连蛋白 (LN)与血管内皮生长因子 (VEGF)表达的意义。方法 :采用免疫组织化学SP法检测 3 4例涎腺腺样囊性癌LN和VEGF的表达。结果 :LN表达与病理分型、淋巴结转移有关 ,与临床分期、发病部位、肿瘤大小无关 ;VEGF表达与病理分型、淋巴结转移、临床分期有关 ,与发病部位、肿瘤大小无关。结论 :LN和VEGF的表达可作为判断涎腺腺样囊性癌生物学行为的指标。     

TGF-β1在口腔颌面部恶性肿瘤中的表达

目的 探讨转化生长因子β1（TGF－β1）在人口腔颌面部恶性肿瘤细胞中的表达水平与其生物学行为的关系。方法 以ELISA法分别检测人口腔颌面部手术切除恶性肿瘤组织细胞，人舌鳞状细胞癌细胞系Tca8113，人涎腺腺样囊性癌细胞系ACC－2和人高转移性涎腺腺样囊性癌细胞系ACC－M，及正常口腔黏膜与黏膜下组织细胞各样本的TGF－β1的表达水平。结果 口腔合面部恶性肿瘤组织TGF－β1的表达水平与正常对照组有非常显著差异（P＜0．01）。Tca8113,ACC-2细胞TGF－β1的表达水平增高正常对照组细胞（P＜0．05）；舌鳞癌系Tca8113与腺要囊性癌细胞系ACC－2的TGF－β1的表达水平无显著差异；高转移腺样囊性癌细胞系ACC－M的TGF－β1的表达水平与ACC－2相比，有非常显著的降低（P＜0．01）；高转移腺样囊性癌细胞系ACC－M无血清培养液中TGF－β1的分泌量亦低于任何其它细胞。结论 TGF－β1的表达水平可能与恶性肿瘤细胞的增殖，侵袭和转移有密切关系。     

蛋白激酶CK2-β在腺样囊性癌细胞中的活性及表达

我们对比了涎腺腺样囊性癌肺高转移细胞(salivary adenoid carcinoma lung metastasis，SACC-LM)与低转移的涎腺腺样囊性癌细胞(SACC-83)中蛋白激酶CK2-β的活性，探讨蛋白激酶CK2-β与涎腺腺样囊性癌发生肺转移的关系。     

涎腺恶性肿瘤病人免疫功能测定与近期预后关系的初步探讨

涎腺恶性肿瘤不仅比较常见,而且复发和全身转移也不少。根据我科的资料统计,涎腺肿瘤为颌面部肿瘤的34.13％,恶性的占涎腺肿瘤的36.1％。上海第二医学院病理科统计涎腺癌约有25％发生淋巴结转移;据其它文献报道腺样囊腺癌区域淋巴结转移高达     

Expression of IgSF in salivary adenoid cystic carcinoma and its relationship with invasion and metastasis

BACKGROUND: The aim of the present work was to study the potential effect of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and neural cell adhesion molecule (NCAM) of immunoglobulin superfamily on the invasion and metastasis of salivary adenoid cystic carcinoma (SACC). METHOD: The expressions of ICAM-1, VCAM-1 and NCAM of forty cases with SACC were examined by immunohistochemical methods using respective kits. RESULTS: A significant relation showed between the expression level and histological differentiation, expression of ICAM-1 and VCAM-1 in solid SACC was greatly increased compared with NCAM; SACC with metastatic lymph node or local recurrence displayed significant relationship to the up-regulation of ICAM-1 and VCAM-1, and to the down-regulation of NCAM. CONCLUSIONS: It is proposed that ICAM-1 and VCAM-1 may play a role in the invasion and metastasis of SACC. NCAM may be an invasion-resistant adhesion molecule.     

不同fimA基因型牙龈卟啉单胞菌对人脐静脉内皮细胞产生血管细胞黏附分子1和细胞间黏附分子1的影响

目的 观察不同fimA基因型牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)产生血管细胞黏附分子1(vascular cell adhesion molecule-1,VCAM-1)和细胞间黏附分子1(intercellular adhesion molecule-1,ICAM-1)的影响,探讨Pg在动脉粥样硬化发生、发展中的可能作用.方法 实验分别以PgATCC33277 (Ⅰ fimA ) 、WCSP115 (Ⅱ fimA)、W83 (Ⅳ fimA)和大肠杆菌脂多糖刺激HUVEC作为T1、T2、T3组(3个实验组)和阳性对照组,未受刺激的HUVEC作为阴性对照组;标准条件下厌氧培养上述3型Pg,将其以及大肠杆菌脂多糖分别与 HUVEC共同孵育2、6、24 h,采用流式细胞术检测HUVEC表面ICAM-1和VCAM-1的蛋白表达量,并通过激光共聚焦显微镜观察ICAM-1和VCAM-1的表达分布情况.结果 Ⅰ、Ⅱ、Ⅳ fimA型Pg刺激HUVEC后,细胞表面ICAM-1表达均增强(P<0.05),2、6、24 h表达量分别为Ⅰ fimA:60.27±5.43、80.81±1.44、85.94±2.56;Ⅱ fimA:86.69±8.81、90.19±0.00、96.18±0.48,Ⅳ fimA:59.66±0.40、85.79±4.86、96.04±2.07.除2 h时ⅠfimA与Ⅳ fimA型Pg刺激的HUVEC表面ICAM-1表达量差异无统计学意义外,其他各时间点Ⅱ、Ⅳ fimA型Pg的刺激作用均强于Ⅰ fimA型Pg(P<0.05).本研究条件下,Ⅰ、Ⅱ、Ⅳ fimA型Pg刺激HUVEC后2、6、24 h表达VCAM-1的水平均较低,各实验组与对照组间相比差异均无统计学意义(P>0.05).激光共聚焦显微镜观察显示,Pg刺激下HUVEC表达ICAM-1和VCAM-1增加,在Ⅱ、Ⅳ fimA型Pg刺激下,HUVEC中ICAM-1和VCAM-1荧光点相对较多且分布范围广.结论 牙周主要致病菌Pg毒力和致病性与其fimA基因型相关,Ⅱ fimA和Ⅳ fimA型Pg 有较强的上调HUVEC表达细胞黏附分子的能力,可能导致血管内皮功能紊乱.

Abstract：

Objective To investigate the effect of Porphyromonas gingivalis(Pg) with different fimA genotypes on vascular cell adhesion molecule-1(VCAM-1) and intercellular adhesion molecule-1(ICAM-1) production by human umbilical vein endothelial cells(HUVEC). Methods In the present study, PgATCC33277(type Ⅰ fimA genotype), WCSP 115(type Ⅱ fimA genotype), W83(type Ⅳ fimA genotype), and Escherichia coli-lipopolysaccharide (Ec-LPS) were designed as experimental group 1, 2, 3, and positive control group, respectively, to stimulate HUVEC, and the un-stimulated HUVEC were analyzed as negative control group. The three strains of Pg were cultured anaerobically in standard condition, and then the Pg cells and Ec-LPS were co-cultured with HUVEC for 2, 6, and 24 h, respectively. The amount of ICAM-1 and VCAM-1 produced by HUVEC was detected with flow cytometry(FCM). The expression of ICAM-1 and VCAM-1 by HUVEC were assayed with confocal laser scanning microscope(CLSM). ResultsThe expression of ICAM-1 on the surface of HUVEC were intensified after infected by Pg with Ⅰ, Ⅱ, and Ⅳ fimA genotypes (P<0.05). The amounts of ICAM-1 were 60.27±5.43, 80.81±1.44, and 85.94±2.56 for Pg with type Ⅰ fimA genotype, 86.69±8.81, 90.19±0.00, and 96.18±0.48 for Pg with type Ⅱ fimA genotype, 59.66±0.40, 85.79±4.86, and 96.04±2.07 for Pg with type Ⅳ fimA genotype at 2, 6 and 24 h after infection, respectively. The up-regulation effects caused by Pg with type Ⅱ and Ⅳ fimA genotypes were stronger than those caused by Pg with type Ⅰ fimA genotype at different time points except at 2 h(P<0.05). Under the present experimental condition, infected by Pg with type Ⅰ, Ⅱ and Ⅳ fimA genotypes stimulated low expression of VCAM-1 by HUVEC, it showed no significant differences among all the groups (P>0.05). Expression of ICAM-1 and VCAM-1 in Pg infected HUVEC were confirmed by CLSM. Infection of HUVEC with Pg resulted in more fluorescence staining of ICAM-1 and VCAM-1 compared with that in uninfected HUVEC cultures. Conclusions The virulence and pathogenicity of Pg is associated with its fimA genotypes, Pg with type Ⅱ and Ⅳ fimA genes possess stronger ability to stimulate HUVEC to up-regulate the expression of cell adhesion molecules, which may lead to disorders in vascular endothelial function.     

Expression of ICAM-1 in implanted primary and metastatic squamous cell carcinomas in rats

FF6 tumor cells are dervied from a spontaneous rat squamous cell carcinoma (SCC) which originally arose in the facial skin of a DA rat. In this study, FF6 tumor cells were implanted into rat oral mucosa to establish an in vivo metastatic model. We analyzed the expression of intercellular cell adhesion molecule-1 (ICAM-1) in the implanted primary and metastatic FF6 tumors by immunostaining with a monoclonal antibody (mAb) against ICAM-1. The implanted primary FF6 cells showed strong expression of ICAM-1, whereas the tumor cells of metastatic lesions showed weak or negative expression of ICAM-1. By imiminostaining with rnAb OX6, a number of MHC class II-positive macro-phages were detected in tumor mesenchyme and surrounding the metastatic foci. These results suggested that the local immune reaction in the lymph node influenced the expression of ICAM-1 on tumor cells, and that MHC class II-positive macrophages may play a role in transplanted tumor growth and metastases.     

Induction of adhesion molecule expression on blood vessels and keratinocytes in recurrent oral ulceration

Adhesion molecules are known to play a crucial role in the recruitment of inflammatory cells to sites of inflammation. In this study endothelial cell and keratinocyte adhesion molecule expression in recurrent oral ulcers (ROU) ( n =13) was compared with that found in normal oral mucosa (NOM) ( n =11) and experimentally induced ulcers (EIU) ( n =5) by using immunohistochemistry. Significantly greater expression of both vascular cell adhesion molecule-1 (VCAM-1) and E-selectin was demonstrated on vasculature in ROU compared with that found in both NOM and EIU. Induction of keratinocyte intercellular adhesion molecule-1 (ICAM-1) was also a prominent feature of ROU. The expression of VCAM-1 and E-selectin on blood vessels in ROU is likely to be important in the accumulation of lymphocytes that characterise early aphthous lesions. The induction of keratinocyte ICAM-1 may facilitate lymphocyte invasion of the epithelium in ROU, which may ultimately result in ulcer formation.     

血管细胞粘附分子-1在口腔鳞状细胞癌中的表达及其与血管生成的关系

目的 观察口腔鳞状细胞癌组织中血管细胞粘附分子-1（vascular cell adhesion molecule-1，VCAM-1）的表达与定位及其与微血管密度（microvessel density，MVD）间的关系，探讨VCAM-1在口腔鳞状细胞癌血管生成中的意义。方法 用免疫组化二步法检测48例口腔鳞状细胞癌组织和10例正常口腔粘膜组织中VCAM-1和CD31表达并计数微血管密度（MVD）。结果 VCAM-1蛋白主要定位于肿瘤细胞的胞膜和胞浆以及血管内皮细胞；VCAM-1在口腔鳞状细胞癌组织中的表达率显著高于正常口腔粘膜组织（P〈0．01）；口腔鳞状细胞癌组织的MVD值显著高于正常口腔粘膜组织（P〈0．01），MVD值与肿瘤浸润深度和有无淋巴结转移密切相关（P〈0．01）；VCAM-1表达阳性组MVD值显著高于VCAM-1表达阴性组（P〈0．01），且VCAM-1表达与MVD呈正相关（P〈0．01）。结论 VCAM-1在口腔鳞状细胞癌组织中的高表达可能在肿瘤的浸润和转移中起重要作用，并与肿瘤血管生成有关。     

The adhesion molecules NCAM, HCAM, PECAM-1 and ICAM-1 in normal salivary gland tissues and salivary gland malignancies

BACKGROUND: Some malignant salivary gland tumors are known for their propensity to exhibit perineural invasion and vascular metastases. It was hypothesized that alterations in the expression of cell adhesion molecules are involved in these processes. METHODS: The expression and distribution of neural cell adhesion molecule (NCAM), HCAM (CD44), platelet-endothelial cell adhesion molecule-1 (PECAM-1), and intercellular cell adhesion molecule-1 (ICAM-1) in normal salivary gland tissues and selected salivary gland malignancies, especially adenoid cystic carcinoma (AdCyCa) and polymorphous low-grade adenocarcinoma (PMLG), were determined immunohistochemically, and their influence on histologically demonstrated perineural invasion, vascular invasion, and tumor recurrence/patient death were investigated. RESULTS: NCAM, HCAM, and ICAM-1 were often found to be expressed by neoplastic cells, but no correlation to perineural invasion, tumor behavior, or patient prognosis was found. PECAM-1 was rarely and only focally expressed in three tumors, all of which were related to tumor metastases and patient death. CONCLUSIONS: Immunohistochemical demonstration of NCAM, HCAM, and ICAM-1 is not related to perineural invasion or tumor behavior. PECAM-1 expression was related to vascular invasion and poor patient prognosis in three cases.     

血管细胞黏附分子-1在口腔鳞状细胞癌中的表达及其意义

目的研究血管细胞黏附分子-1（VCAM-1）在口腔鳞状细胞癌（OSCC）中的表达与定位及其临床意义。方法应用分子原位杂交和免疫组化方法对48例OSCC组织和10例正常口腔黏膜组织中VCAM-1 mRNA和VCAM-1蛋白质的表达和定位进行检测,比较VCAM-1在不同口腔组织中的表达率及其与临床指标的关系。结果VCAM-1蛋白定位于肿瘤细胞胞浆和胞膜,VCAM-1 mRNA定位于肿瘤细胞胞浆。VCAM-1 mRNA和VCAM-1蛋白在OSCC中的表达率均显著高于正常口腔黏膜组织（P＜0.01）,VCAM-1 mRNA表达与VCAM-1蛋白表达呈正相关（P＜0.01）;OSCC中淋巴结转移者VCAM-1蛋白的表达显著高于无淋巴结转移者（P＜0.01）。结论OSCC中VCAM-1基因的高表达可能与肿瘤的浸润和转移有关。     

血管生成相关因子在口腔扁平苔藓中的表达

目的 研究血管生成相关因子血管内皮生长因子(vascular endothelial growth factor,VEGF)、细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)、血管细胞黏附分子-1(vascular cell adhesion molecule-1,VCAM-1)在口腔扁平苔藓患者血清和组织中的表达情况.方法 30例扁平苔藓患者纳入扁平苔藓组,15例健康志愿者作为对照(正常组)用于血清收集,收集扁平苔藓组织患者活检组织30例及外科手术切除的多余正常口腔黏膜组织15例,应用免疫组织化学技术分析VEGF、ICAM-1、VCAM-1分别在扁平苔藓病损局部组织及正常黏膜组织中的表达情况.结果 ELISA结果显示VEGF在2组血清中表达差异无统计学意义(t=1.837,P=0.074),而ICAM-1(t=2.730,P=0.009)、VCAM-1(t=2.167,P=0.035)在扁平苔藓组血清中的表达明显高于正常组.免疫组织化学染色结果显示VEGF、ICAM-1、VCAM-1在口腔黏膜正常上皮中呈现少量阳性表达或阴性表达,而在扁平苔藓组织中VEGF(x2=33.632,P<0.05)、ICAM-1(x2=45.000,P<0.05)、VCAM-1(x2=37.286,P<0.05)表达较正常黏膜组织均出现上调趋势.结论 在扁平苔藓患者的血清和病变组织中,血管生成相关因子表达上调.     

Differential expression of interleukin-8 and intercellular adhesion molecule-1 by human gingival epithelial cells in response to Actinobacillus actinomycetemcomitans or Porphyromonas gingivalis infection

Little is known regarding the molecules expressed by gingival epithelial cells that are involved in initiating and maintaining inflammation following the interaction with periodontal pathogens. Thus, we investigated the effect of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis infection on the expression of neutrophil chemoattractant interleukin 8 (IL-8) and the adhesion molecule intercellular adhesion molecule-1 by gingival epithelial cells. The data revealed that both IL-8 and intercellular adhesion molecule-1 expression increased after infection with A. actinomycetemcomitans (IL-8: 2- to 7-fold; intercellular adhesior molecule-1: 2.5- to 3.7-fold). IL-8 secretion reached a maximal level 6 h after the infection and the expression subsequently decreased to basal level. The increased cell surface intercellular adhesion molecule-1 expression started at 4 h after infection and reached a maximal level 14 h after the infection. In contrast, the expression of both molecules rapidly decreased 2 h after challenge with P. gingivalis. This opposite influence of A. actinomycetemcomitans and P. gingivalis infection on the expression of IL-8 and intercellular adhesion molecule-1 by gingival epithelial cells suggests that A. actinomycetemcomitans infection may initiate the recruitment of neutrophils, whereas the P. gingivalis infection may retard this process and therefore demonstrate a distinct perspective of virulence.     

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.