MACC1和C-Met在口腔白斑和口腔鳞状细胞癌中的表达及意义

目的:探讨分析MACC1和C-Met在正常口腔黏膜、口腔白斑及口腔鳞状细胞癌中的表达及其临床意义。方法:采用免疫组化SP法检测20例口腔黏膜、20例上皮异常增生白斑、50例口腔鳞癌组织中的MACC1、C-Met蛋白的表达情况,采用X2和spearman等级相关分析对结果进行判定。结果:MACC1、C-Met蛋白在异常增生型白斑和口腔鳞癌中的阳性表达率分别为50%、76%,35%、66%,均明显高于正常口腔黏膜(17.6%,5.0%),差异均有统计学意义(P0.05)。MACC1和C-Met蛋白表达与口腔鳞癌的分期、淋巴结转移及分化程度密切相关(P0.05)。Spearman等级相关分析显示口腔白斑及口腔鳞癌中MACC1和C-Met的表达呈现正相关(P0.05)。结论:MACC1和C-Met在上皮不典型增生性白斑和口腔鳞癌中高表达,二者在口腔黏膜白斑的癌变和口腔鳞癌的发生发展中可能起重要作用。     

PLCE1与MMP-9在口腔鳞癌中的表达及其临床意义*

目的:探讨磷脂酶Cε1(PLCE1)与基质金属蛋白酶-9(MMP-9)在口腔鳞癌中的表达及临床意义。方法:采用免疫组化方法检测61例口腔鳞癌组织和35例正常口腔黏膜组织中PLCE1和MMP-9的蛋白表达,并分析二者与口腔鳞状细胞癌临床病理参数的关系及二者的相关性。结果:PLCE1在口腔鳞癌组织中表达阳性率为68.85%(42/61),MMP-9在口腔鳞癌组织中表达阳性率为75.40%(46/61),两者在正常口腔黏膜组织中表达阳性率分别为14.28%(5/35)、17.14%(6/35)。PLCE1和MMP-9在正常口腔黏膜组织的阳性表达率均明显低于口腔鳞癌组织(P0.01)。PLCE1与MMP-9的高度阳性表达和患者的性别、年龄、吸烟及肿瘤大小无明显相关性,但与肿瘤TNM分期以及组织分化程度显著相关。口腔鳞癌组织中PLCE1和MMP-9的高表达呈明显正相关(r=0.438,P0.01)。结论:PLCE1和MMP-9的过度表达均与口腔鳞癌的发生、发展及侵袭转移有密切相关性,并检测二者可能会对口腔鳞癌患者的早期临床诊断及预后判断具有一定的参考价值。     

PD-L1和CD147在口腔鳞癌中的表达与临床意义

目的:探讨口腔鳞癌组织中免疫共刺激分子PD-L1与细胞外基质蛋白酶诱导因子CD147的表达、两者的相关性及临床意义。方法:应用免疫组化技术检测66例口腔鳞癌组织及36例正常口腔黏膜组织中PD-L1和CD147的表达,分析PD-L1、CD147表达的相关性及二者与口腔鳞癌临床病理参数的关系。结果:PD-L1在口腔鳞癌组织中表达阳性率为68.18%(45/66),正常口腔黏膜组织中表达阳性率仅为16.67%(6/36);CD147在口腔鳞癌组织中表达阳性率为74.24%(49/66),明显高于其在正常口腔黏膜组织中的表达13.88%(5/36)。PD-L1和CD147两者在口腔鳞癌组织中阳性表达率与口腔黏膜组织相比均明显升高(P0.01)。统计学分析显示,PD-L1和CD147在口腔鳞癌组织中的高表达与患者的性别年龄、吸烟史及肿瘤的体积等因素无明显相关,但与TNM分期及鳞癌的组织分化程度紧密相关。口腔鳞癌组织中PD-L1与CD147两者相关性分析r=0.342,P值小于0.01,说明二者的表达呈显著正相关。结论:口腔鳞癌组织中PD-L1与CD147均呈高表达,并且二者的过度表达可能与口腔鳞癌的发生、发展关系密切,合并检测二者可能为OSCC的诊疗及预后指明新的方向,为口腔鳞癌的靶向治疗提供新的靶点。     

AEG-1和MMP-9在口腔鳞癌中的表达与临床意义

目的:研究星形细胞提升基因-1(AEG-1)与基质金属蛋白酶-9(MMP-9)在口腔鳞癌组织中的表达及其临床意义。方法:采用免疫组化的方法检测53例口腔鳞癌组织和30例正常口腔黏膜组织中AEG-1和MMP-9的蛋白表达,分析其与口腔鳞癌临床病理参数的关系及二者的相关性。结果:AEG-1和MMP-9在口腔鳞癌组织中表达阳性率分别为66.03%(35/53)、73.58%(39/53),在正常口腔黏膜组织中表达阳性率分别为13.33%(4/30)、16.67%(5/30)。AEG-1和MMP-9在口腔鳞癌组织中阳性表达率均高于正常口腔黏膜组织(P0.01)。高表达的AEG-1和MMP-9与口腔鳞癌的组织分化程度,淋巴结转移以及临床分期有密切相关性,但与患者的性别、年龄及吸烟史无相关性。口腔鳞癌组织中AEG-1与MMP-9的表达呈显著正相关(r=0.474,P0.01)。结论:AEG-1和MMP-9的高表达均与口腔鳞癌的发生、发展及转移有密切相关性,同时检测二者可能会对口腔鳞癌的早期诊断及预后判断具有一定的临床参考价值。     

口腔白斑抑癌基因DAPK和TIG1高甲基化表达研究

摘要 目的：探讨抑癌基因DAPK、TIG1高甲基化在口腔白斑中表达状态及其对口腔癌发生发展中的作用。方法：取77例口腔白斑、32例口腔鳞癌、32份正常口腔黏膜组织,用实时定量甲基化特异性PCR技术检测组织中DAPK、TIG1高甲基化表达并进行统计学分析。结果：DAPK在口腔鳞癌组织中高甲基化表达率为46.9%,表达量为（0.0728±0.1617）,明显高于其在口腔白斑组织（19.5%,0.0070±0.0172）和口腔正常组织（18.8%,0.0021±0.0050）中的表达,差异有统计学意义（P<0.05）。DAPK高甲基化表达与口腔白斑组织上皮异常增生程度相关,上皮增生高风险组相对于低风险组DAPK高甲基化表达风险增加（OR,1.013;95% CI,1.004-1.023;P=0.004）。TIG1高甲基化在正常组织中未表达,在口腔鳞癌组织和口腔白斑组织表达为（28.1%,0.0174±0.0440）和（27.3%,0.0035±0.0076）,与正常组织相比具有统计学意义（P<0.05）。结论：抑癌基因 DAPK、TIG1高甲基化有望成为口腔黏膜癌变早期标志物。     

口腔疣状癌和口腔鳞癌组织中E-cadherin表达的对比研究

目的:研究E-钙粘素(E-cadherin,E-cd)在口腔疣状癌和口腔鳞癌中的表达,探讨E-cd与口腔疣状癌,口腔鳞癌分化程度的关系。方法:采用免疫组织化学和免疫电镜方法检测18例口腔疣状癌、20例口腔鳞癌组织中的E-cd表达。结果:E-cd在正常口腔黏膜、口腔疣状癌、高分化鳞癌和低分化鳞癌中主要位于胞膜及胞浆上。电镜观察显示E-cd在胞膜主要定位于桥粒,胞浆主要定位于粗面内质网和线粒体上。定量检测显示:E-cd在正常口腔黏膜、口腔疣状癌、高分化鳞癌和低分化鳞癌中的阳性表达率分别100%(8/8)、61.1%(11/18)、50.0%(5/10)和30.0%(3/10)。口腔疣状癌E-cd蛋白的平均染色强度与高分化鳞癌间差异无显著性,但明显低于正常黏膜组(P<0.05)而高于低分化鳞癌(P<0.05)。结论:E-cd的表达水平随口腔肿瘤分化程度的降低而降低,且超微结构的表达部位与肿瘤的分化程度存在相关性。     

人口腔鳞癌组织中HIF-1a和NF-kB的表达及意义

目的:探讨缺氧诱导因子(HIF-1a)和核因子-KB(NF-KB)口腔鳞癌中的表达及相互关系,研究它们的表达与肿瘤临床病理指标的联系.方法:应用SP染色法检测HIF-1a和NF-KB在49例口腔鳞癌组织、10例正常口腔黏膜组织中的阳性率.结果在口腔鳞癌中HIF-1a和NF-KB的阳性表达率分别为80.0%和78.4%,其阳性率及表达等级均显著高于正常对照组(P<0.05),在中一低分化组和有淋巴结转移组中的表达显著高于高分化组和无淋巴结转移组(P<0.05).HIF-1a表达与NF-KB表达成等级正相关(r=0.45,P<0.05).结论:HIF-1a或NF-KB与口腔鳞癌生物学行为有密切关系,二者的联合检测,有助于口腔鳞癌恶性程度和生物学特性的判断.     

DKK-1和MMP-14在口腔鳞癌中的表达及其意义

目的:研究DKK-1蛋白和MMP-14蛋白在口腔鳞状细胞癌组织中的表达和临床意义。方法:选择口腔鳞癌石蜡标本62例作为实验组,25例正常口腔黏膜组织作为对照组。应用免疫组织化学法检测其DKK-1和MMP-14蛋白的表达,并分析二者与口腔鳞癌患者临床病理特征之间的关系及二者表达的相关性。结果:实验组DKK-1的阳性表达率为40.32%,明显低于对照组中(68%),而MMP-14的阳性表达率(72.58%)明显高于对照组(24%)(P0.05)。DKK-1和MMP-14的表达水平与口腔鳞癌的分期、淋巴结转移及分化程度有显著相关(P0.05)。口腔鳞癌组织中DKK-1和MMP-14蛋白的表达呈显著负相关(r=-0.600,P0.05)。结论:口腔鳞状细胞癌组织中DKK-1的表达下调,MMP-14的表达上调,二者可能参与了OSCC的发生和发展过程,并有望用于口腔鳞状细胞癌的病情和预后评估。     

Anxa2, Stat3 在口腔鳞状细胞癌中的表达及意义

目的:探讨膜联蛋白A2(Anxa2)和细胞信号转导和转录激活因子3(Stat3)在口腔鳞癌组织中的表达及其临床意义。方法:选择口腔鳞癌石蜡标本80例为实验组,20例正常口腔黏膜组织为对照组。应用免疫组织化学法检测Anxa2和Stat3蛋白的阳性表达,并进行结果判定,采用x2和Spearman等级相关分析法分析二者表达差异性及其相关性。结果:Anxa2、Stat3在病例组中的阳性表达率,分别为81.3%(65/80)、87.5%(70/80),明显高于对照组中的表达率,25.0%(5/20)、30.0%(6/20),且差异有统计学意义(P0.05),Anxa2和Stat3蛋白表达水平与口腔鳞癌的分期、淋巴结转移及分化程度有密切关系。Spearman等级相关分析Anxa2和Stat3在口腔鳞癌组织中蛋白的表达呈正相关(r=0.302,P0.01)。结论:Anxa2、Stat3在OSCC的发生和转移过程中均具有重要的作用,且二者间也有相互作用的关系。     

口腔鳞癌组织细胞凋亡相关蛋白Fas/FasL的表达研究

目的　研究调亡相关蛋白Fas、FasL在口腔鳞状细胞癌组织中的表达及意义。方法　应用免疫组织化学方法检测 10例正常口腔粘膜、 38例口腔鳞癌组织及肿瘤浸润淋巴细胞 (TIL)和 11例转移淋巴结中Fas、FasL的表达。结果　Fas在正常口腔粘膜中广泛表达 ;鳞癌组织表达明显下调 (P <0 .0 5 ) ;Fas表达与口腔鳞癌分化程度有关 ;淋巴结转移癌中Fas表达减弱。FasL在正常口腔粘膜不表达 ;鳞癌组织表达明显上调 (P <0 .0 5 ) ;FasL表达与口腔鳞癌分化程度无关 (P >0 .0 5 ) ;有淋巴结转移者FasL阳性表达率高于无转移者 (P <0 .0 5 )。TIL细胞Fas、FasL阳性表达率为 81.6 %和 84 .2 % .。Fas、FasL在口腔鳞癌的表达有明显相关性 (P <0 .0 5 )。结论　Fas表达与口腔粘膜上皮细胞的自然分化成熟衰老及口腔鳞癌的形成和肿瘤的恶性度有关。FasL的表达上调可能是口腔鳞癌组织免疫反攻击的体现 ,对促进肿瘤的发生发展及转移有重要作用。     

Analysis of silver binding nucleolar organizer regions in exfoliative cytology smears of potentially malignant and malignant oral lesions

Nucleolar organizer regions are nucleolar components that contain proteins that are stained selectively by silver methods; they can be identified as black dots throughout the nucleolus and are known as silver binding nucleolar organizer regions (AgNOR). The number of AgNOR is related to the cell cycle and the proliferative activity of the cells. We investigated AgNOR using exfoliative cytology smears of potentially malignant oral lesions. Eighty individuals were divided into four equal groups: healthy controls, oral leukoplakia, oral submucous fibrosis and oral squamous cell carcinoma. The mean number of AgNOR in each study group gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. The proliferative index was increased in the oral premalignant and malignant patients compared to normal subjects. The mean AgNOR size gradually increased from control to oral leukoplakia to oral submucous fibrosis to oral squamous cell carcinoma. Spherical shaped AgNOR were most common in controls, whereas large, clustered and kidney shapes were most common in oral squamous cell carcinoma. Multiparameter analysis of AgNOR in oral exfoliative smears is a simple, sensitive and cost-effective method for differentiating premalignant from malignant lesions and can be used in conjunction with routine cytomorphological evaluation.     

Computer aided morphometric analysis of oral leukoplakia and oral squamous cell carcinoma

We compared the changes in the cells in the basal layer of normal mucosa, oral leukoplakia with dysplasia and different grades of oral squamous cell carcinoma (OSCC) using computer aided image analysis of tissue sections. We investigated three morphometric parameters: nuclear area (NA), cell area (CA) and their ratio (NA:CA). NA and NA:CA ratio showed a statistically significant increase from dysplasia to increasing grades of OSCC. Nuclear size was useful for differentiating normal tissue, potentially malignant leukoplakia and OSCC.     

口腔白斑组织癌变标志性基因筛查

口腔黏膜白斑病（oral leukoplakia,OLK）是最常见的口腔粘膜的癌前病变．OLK恶性转化率高达43%,因此加强其风险调查和监测对于该病的预防和治疗具有重要意义．本研究采用美国SuperArray公司的肿瘤基因芯片（OHS-802）检测口腔正常组织、OLK组织和口腔鳞状细胞癌（oral squamous cell carcinoma, OSCC)组织的肿瘤相关基因表达差异,旨在筛查与口腔黏膜白斑病发生和癌变相关的表达阳性基因,为探究OLK的发生及癌变机制奠定基础．将“正常组织→OLK组织→OSCC组织”连续递增2倍以上或递减2倍以上的基因确定为OLK组织癌变的标志性基因．SuperArray基因芯片、RT-PCR法和实时定量PCR结果显示,仅有3个基因（ACP-2、BCL-2、SOCS-3）表达水平连续下调2倍以上,有6个基因（CLK-3、CTNNB-1、FKBP-8、GDF-15、NF-1、XRCC-1）表达连续上调2倍以上．进一步经RT-PCR和实时定量PCR验证,其结果与SuperArray结果一致．本研究结果提示,ACP-2等9个基因可能是口腔正常黏膜组织经OLK转变为癌组织的驱动基因,可作为OLK组织、OSCC组织和口腔正常黏膜组织诊断的分子标志．     

原发性口腔鳞癌组织及血清中内皮抑素表达的意义

目的：探讨原发性口腔鳞癌患者组织和血清中内皮抑素表达及与肿瘤分期、分级的关系。方法：采用免疫组化方法检测36例口腔鳞癌和12例正常口腔粘膜组织中内皮抑素表达情况。ELISA法检测36例口腔鳞癌患者术前血清内皮抑素水平,14例健康者血清做对照。结果：内皮抑素主要见于肿瘤组织细胞质。正常口腔粘膜中内皮抑素表达率为7.15%,口腔鳞癌组织中内皮抑素阳性率为76.44%,其中G1、G2、G3级阳性率分别为47.21%、79.17%、90.90%,病理分级间比较差异有统计学意义（P〈0.05）。口腔鳞癌患者血清中内皮抑素水平（49.62±1.72）ng/mL显著高于健康对照者（5.60±0.37）ng/mL（P〈0.05）,TNM分期III、IV期肿瘤患者血清内皮抑素水平显著高于I和II期（P〈0.05）。结论：口腔鳞癌患者组织和血清中内皮抑素表达显著升高,并与肿瘤分期、分级相关,检测内皮抑素表达有助于判断口腔鳞癌恶性程度。     

miR-146a Enhances the Oncogenicity of Oral Carcinoma by Concomitant Targeting of the IRAK1, TRAF6 and NUMB Genes

MicroRNAs are short non-coding RNAs that regulate gene expression and are crucial to tumorigenesis. Oral squamous cell carcinoma (OSCC) is a prevalent malignancy worldwide. Up-regulation of miR-146 has been identified in OSCC tissues. However, the roles of miR-146 in carcinogenesis are controversial as it is suppressive in many other malignancies. The present study investigated the pathogenic implications of miR-146a in oral carcinogenesis. Microdissected OSCC exhibits higher levels of miR-146a expression than matched adjacent mucosal cells. The plasma miR-146a levels of patients are significantly higher than those of control subjects; these levels decrease drastically after tumor resection. miR-146a levels in tumors and in patients’ plasma can be used to classify OSCC and non-disease status (sensitivity: >0.72). Exogenous miR-146a expression is significantly increased in vitro oncogenic phenotypes as well as during xenograft tumorigenesis and OSCC metastasis. The plasma miR-146a levels of these mice parallel the xenograft tumor burdens of the mice. A miR-146a blocker abrogates the growth of xenograft tumors. miR-146a oncogenic activity is associated with down-regulation of IRAK1, TRAF6 and NUMB expression. Furthermore, miR-146a directly targets the 3′UTR of NUMB and a region within the NUMB coding sequence when suppressing NUMB expression. Exogenous NUMB expression attenuates OSCC oncogenicity. Double knockdown of IRAK1 and TRAF6, and of TRAF6 and NUMB, enhance the oncogenic phenotypes of OSCC cells. Oncogenic enhancement modulated by miR-146a expression is attenuated by exogenous IRAK1 or NUMB expression. This study shows that miR-146a expression contributes to oral carcinogenesis by targeting the IRAK1, TRAF6 and NUMB genes.     

Salivary mRNA markers having the potential to detect oral squamous cell carcinoma segregated from oral leukoplakia with dysplasia

BackgroundIn the current study the presence of extracellular IL-1B, IL-8, OAZ and SAT mRNAs in the saliva was evaluated as a tool in the early detection of oral squamous cell carcinoma.Methods34 patients with primary oral squamous cell carcinoma stage T₁N₀M₀/T₂N₀M₀, 20 patients with oral leukoplakia and dysplasia (15 patients with mild dysplasia and 5 with severe dysplasia/in situ carcinoma) and 31 matched healthy-control subjects were included in the study. The presence of IL-1B, IL-8, OAZ and SAT mRNA was evaluated in extracellular RNA isolated from saliva samples using sequence-specific primers and real-time RT-PCR. ROC curve analysis was used to estimate the ability of the biomarkers to detect oral squamous cell carcinoma patients.ResultsThe data reveal that the combination of these four biomarkers provides a good predictive probability of up to 80% (AUC = 0.799, p = 0.002) for patients with oral squamous cell carcinoma but not patients suffering from oral leukoplakia with dysplasia. Moreover, the combination of only the two biomarkers (SAT and IL-8) also raises a high predictive ability of 75.5% (AUC = 0.755, p = 0.007) approximately equal to the four biomarkers suggesting the use of the two biomarkers only in the prediction model for oral squamous cell carcinoma patients limiting the economic and health cost in half.ConclusionSAT and IL-8 mRNAs are present in the saliva in high quality and quantity, with a good discriminatory ability for oral squamous cell carcinoma patients only but not for patients with oral leukoplakia and dysplasia an oral potentially malignant disorder.     

The Combination of SMAD4 Expression and Histological Grade of Dysplasia Is a Better Predictor for the Malignant Transformation of Oral Leukoplakia

Oral leukoplakia (OL) is the most common premalignancy in the oral cavity and can progress to oral squamous cell carcinoma (OSCC). SMAD4 is a tumor suppressor implicated in multiple cancer types including OSCC. To assess the role of SMAD4 in oral leukoplakia malignant transformation, the authors investigated SMAD4 expression patterns in OL and OSCC using a highly specific antibody and correlated the patterns with the risk of malignant transformation oral leukoplakia. Immunohistochemistry and a quantitative imaging system were used to measure SMAD4 expression in OL from 88 OL patients, including 22 who later went through malignant transformation, and their OSCC counterpart. Forty-three (48.9%) of the 88 OL patients had strong SMAD4 expression. SMAD4 expression had no significant correlation with patients'' clinicopathological parameters. Interestingly, 17 (39.5%) of the 43 OL lesions with strong SMAD4 expression went through malignant transformation whereas only 5 (11.1%) of the 45 OL lesions with weak SMAD4 expression did so (p = 0.002). The SMAD4 expression in OL was much higher than that in their OSCC counterpart. Kaplan-Meier analysis revealed that the combination of SMAD4 expression and histological grade of dysplasia (p = 0.007) is a better predictor for the malignant transformation of oral leukoplakia. In the multivariate analysis, both SMAD4 expression and grade of dysplasia were identified as independent factors for OL malignant transformation risk (p = 0.013 and 0.021, respectively). It was concluded that high SMAD4 expression may be indicative of an early carcinogenic process in OL and serve as an independent biomarker in assessing malignant transformation risk in patients with OL, and the combination of SMAD4 expression and histological grade of dysplasia is a better predictor for the malignant transformation of oral leukoplakia.     

Improvement in the risk assessment of oral leukoplakia through morphology-related copy number analysis

Oral leukoplakia is the most common type of oral potentially malignant disorders and considered a precursor lesion to oral squamous cell carcinoma. However, a predictor of oral leukoplakia prognosis has not yet been identified. We investigated whether copy number alteration patterns may effectively predict the prognostic outcomes of oral leukoplakia using routinely processed paraffin sections. Comparison of copy number alteration patterns between oral leukoplakia with hyperplasia(HOL,n=22) and dysplasia(DOL, n=21) showed that oral leukoplakia with dysplasia had a higher copy number alteration rate(86%)than oral leukoplakia with hyperplasia(46%). Oral leukoplakia with dysplasia exhibited a wider range of genomic variations across all chromosomes compared with oral leukoplakia with hyperplasia. We also examined a retrospective cohort of 477 patients with oral leukoplakia with hyperplasia with detailed follow-up information. The malignant transformation(MT, n=19)and leukoplakia recurrence(LR, n=253) groups had higher frequencies of aneuploidy events and copy number loss rate than the free of disease(FD, n=205) group. Together, our results revealed the association between the degree of copy number alterations and the histological grade of oral leukoplakia and demonstrated that copy number alteration may be effective for prognosis prediction in oral leukoplakia patients with hyperplasia.