LC3和beclinl在口腔黏膜癌变过程中不同时期的变化及其意义

目的检测微管相关蛋白1轻链3(microtubule-assaiated protein 1 light chain 3,LC3)和自噬基因beclin1在口腔黏膜癌变过程不同时期的特点及表达趋势,探讨其在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)发生进展中的意义。方法采用免疫组织化学方法对19例口腔正常黏膜、9例单纯增生颊黏膜、14例异常增生颊黏膜、15例原位癌阶段颊黏膜及38例OSCC标本进行LC3及beclin1检测。结果 OSCC组LC3和beclin1阳性率显著低于正常口腔黏膜组、单纯增生组、异常增生组及原位癌组;原位癌组LC3和beclin1阳性率较上皮异常增生组虽有升高但无明显差异;LC3和beclin1在OSCC中的表达无相关性。结论 LC3和beclin1在口腔黏膜癌变的过程中表达下调,在原位癌阶段自噬活性有所增加,癌变完成后自噬能力显著降低;自噬活性的改变可能与口腔鳞癌的发生、发展相关。     

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。     

中国地鼠口腔颊囊黏膜癌变过程凋亡相关基因caspase-3、caspase-9、Bax、Bcl-2的表达

目的探讨凋亡相关基因caspase-3、caspase-9、Bax、Bcl-2在中国地鼠口腔正常黏膜、上皮单纯增生、上皮异常增生和鳞状细胞癌组织中的表达及其意义。方法采用免疫组织化学法、RT-PCR检测正常中国地鼠口腔黏膜上皮、单纯增生上皮、异常增生上皮和鳞癌组织中caspase-3、caspase-9、Bax、Bcl-2蛋白及mRNA的表达。结果在口腔黏膜癌变过程中,鳞癌组织中抑制凋亡蛋白Bcl-2表达明显高于口腔正常黏膜、上皮单纯增生、上皮异常增生(P0.05);异常增生上皮caspase-3、caspase-9、Bax表达均明显高于正常组(P0.05),但随着增生程度的加重,caspase-3、caspase-9、Bax表达明显降低(P0.05)。相关性分析显示,鳞癌组织中Bcl-2与Bax、caspase-3、caspase-9呈负相关(P0.05)。RT-PCR结果表明,与正常组织相比,鳞癌组织中Bcl-2高表达,而caspase-3、caspase-9、Bax的mRNA表达显著下调(P0.05)。结论本实验在中国地鼠鳞癌组织中caspase-3、caspase-9、Bax表达降低而Bcl-2表达上升,揭示了caspase-3、caspase-9、Bax、Bcl-2表达与口腔鳞癌的发生、发展密切相关,可以为口腔颊囊黏膜癌的基因治疗提供一些线索或并为评价OSCC的生物学特征及预后提供参考。     

CXXC4、EZH2在口腔鳞状细胞癌中表达及临床意义

目的:探讨口腔鳞状细胞癌(Oral Squamous Cell Carcinoma,OSCC)中CXXC指蛋白4(CXXC finger protein 4,CXXC4)与Zeste基因增强子同源物2(Enhancer of zeste homolog 2,EZH2)的表达、相关性及与临床病理特征之间的关系,为OSCC的早期诊断、治疗、判断预后及预防等提供参考依据。方法:采用免疫组织化学S-P法检测CXXC4蛋白及EZH2蛋白在60例口腔鳞状细胞癌组织及20例非肿瘤患者正常口腔黏膜组织中的表达情况,分析二者与口腔鳞状细胞癌临床病理特征的相关性。结果:口腔鳞状细胞癌组织中CXXC4蛋白的表达率为35.0%(21/60),明显低于正常口腔黏膜组织的60.0%(12/20)(P0.05);口腔鳞状细胞癌中EZH2蛋白的表达率为70.0%(42/60),高于正常口腔黏膜组织的25%(5/20)(P0.05)。CXXC4蛋白与EZH2蛋白的表达与口腔鳞状细胞癌的病理分化程度、TNM分期及淋巴结转移等密切相关(P0.05),而与年龄、性别及肿瘤直径大小等无关。CXXC4蛋白与EZH2蛋白在OSCC中表达呈负相关(r=-0.511,P0.001)。结论:CXXC4蛋白的表达下调或缺失以及EZH2蛋白的表达上调或过表达与口腔鳞状细胞癌的发生、发展及侵袭转移有关,可能作为口腔鳞状细胞癌预后预测的参考指标。     

ING4和HIF-1α在口腔鳞癌中的表达及临床意义

目的:探讨人口腔鳞状细胞癌(OSCC)组织中生长抑制因子4(ING4)和缺氧诱导因子-1α(HIF-1α)的表达及其临床意义。方法:选择我院确诊的口腔鳞癌患者共计65例,均经过手术治疗,术前未行放化疗,切除标本经过石蜡包埋切片,同时选取20例正常口腔黏膜作为对照。采用免疫组化S-P法检测人口腔鳞状细胞癌和正常口腔黏膜组织中ING4和HIF-1α的表达,并分析其与OSCC患者临床病理特征的相关性及人口腔鳞状细胞癌组织中ING4和HIF-1α表达的相关性。结果:OSCC组织中ING4的阳性表达率为41.5%(27/65),显著低于正常口腔黏膜组织(P0.05);OSCC组织中HIF-1α的阳性表达率为64.6%(42/65),显著高于正常口腔黏膜组织(P0.05)。ING4和HIF-1α的表达与OSCC的病理分级、TNM分期和是否有淋巴结转移显著相关(P0.05);OSCC组织中,ING4的表达与HIF-1α的表达呈显著负相关(P0.05)。结论:口腔鳞状细胞癌组织中ING4的表达下调,HIF-1α的表达上调,二者可能通过负向调控作用在口腔鳞状细胞癌的发生、发展、侵袭和转移中发挥着重要作用。     

酪氨酸磷酸酶Z1型受体、环氧合酶-2、人端粒酶逆转录酶在口腔黏膜下纤维性变癌变中的表达及相关性研究

摘要 目的：探究酪氨酸磷酸酶Z1型受体（PTPRZ1）、环氧合酶-2（COX-2）、人端粒酶逆转录酶（hTERT）在口腔黏膜下纤维性变（oral submucosal fibrosis, OSF）癌变中的表达及相关性。方法：采集2018年6月-2019年6月于我院接受治疗的OSF癌变患者OSF癌变组织30例,作为本文研究OSF癌变组;采集2018年6月-2019年6月于我院接受治疗的OSF患者OSF组织30例,作为本文研究OSF组;同时选取无槟榔、烟酒嗜好、正常口腔粘膜组织30例作为本文研究正常组。Western blot法检测PTPRZ1、hTERT相对表达量,酶联免疫吸附法检测COX-2表达,对待测标本进行免疫组化染色,并对PTPRZ1、COX-2、hTERT在OSF癌变中的相关性进行分析。结果：OSF组PTPRZ1、COX-2、hTERT相对表达量均高于正常组（P＜0.05）;OSF癌变组PTPRZ1、COX-2、hTERT相对表达量均高于正常组、OSF组（P＜0.05）。低分化OSF癌变组织中PTPRZ1、COX-2、hTERT相对表达量均高于高分化、中分化OSF癌变组织（P＜0.05）。Ⅳ期OSF癌变组织中PTPRZ1、COX-2、hTERT相对表达量均高于Ⅱ期、Ⅲ期OSF癌变组织（P＜0.05）。OSF癌变组织中PTPRZ1、COX-2、hTERT相对表达量呈正相关。结论：PTPRZ1、COX-2、hTERT在OSF癌变中呈异常高表达,低分化、病理分期较高的OSF癌变中PTPRZ1、COX-2、hTERT表达较高,并且PTPRZ1、COX-2、hTERT在OSF癌变中表达呈正相关。     

NUMB在口腔白斑和鳞癌中的表达变化及意义

目的:研究NUMB在口腔白斑和鳞癌中的表达及意义。方法:采用免疫组织化学法检测88例口腔正常黏膜、口腔白斑及口腔鳞癌石蜡包埋组织中NUMB蛋白的表达。结果:NUMB在口腔正常黏膜(95.7%)、口腔白斑(75%)及口腔鳞癌(4.7%)中均有阳性表达但是表达频率依次降低。NUMB在各个组中的阳性表达率具有统计学差异(P0.05)。结论NUMB阳性表达率随口腔组织恶性程度增高而减少的趋势提示该基因可能在口腔正常黏膜到口腔白斑再到口腔鳞癌的转化中起作用,NUMB有可能成为早期发现癌变的分子生物学标志之一。     

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。     

口腔鳞状细胞癌中hMSH2、PCNA和p53的表达及意义

目的:探讨hMSH2、PCNA和p53在OSCC中的表达及可能存在的临床意义.方法:运用免疫组织化学S-P法对56例OSCC组织中hMSH2、PCNA和p53的表达进行检测.结果:(1)3种基因产物在OSCC中的阳性率均高于正常口腔黏膜,中-低分化癌的阳性率高于高分化癌,有淋巴结转移的阳性率高于无淋巴结转移.(2)hMSH2与PCNA、p53,p53与PCNA的表达均呈正相关.(3)5年生存率与hMSH2蛋白表达和淋巴结转移有关.结论:hMSH2、PCNA和p53的异常表达及其相互之间的调节可能与OSCC的发生发展有关.     

HLA-B在口腔鳞状细胞癌中的表达及意义

目的通过检测人类白细胞抗原B(human leukocyte antigen B,HLA-B)在口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)原发灶、转移灶以及口腔正常黏膜、癌前病变中的表达情况,探讨其在口腔鳞癌发生、发展和淋巴道转移中的作用。方法应用免疫组织化学方法检测70例OSCC组织、14例癌前病变组织以及10例正常组织的HLA-B表达,比较OSCC的不同病理分级、TNM分期、有无淋巴结转移、原发灶和转移灶、正常及癌前病变组织HLA-B的表达情况。结果 HLA-B在正常组织、癌前病变组织、癌组织的表达逐渐下降(P0.05);其中,癌分化等级越低,HLA-B表达量越低;正常组织、癌前病变组织和高分化OSCC之间的表达无明显差异,但明显高于中、低分化OSCC(P0.05);不同T分期之间差异无统计学意义(P0.05);原发灶与转移灶HLA-B表达差异亦无统计学意义(P0.05)。但是,未出现转移的OSCC原发灶HLA-B表达高于出现转移的原发灶(P=0.069)。结论中、低分化OSCC组织HLA-B表达较正常组织、癌前病变以及高分化OSCC明显下调,与中、低分化OSCC较高的淋巴道转移率相关,提示HLA-B的表达减弱可能通过肿瘤免疫逃逸机制在OSCC淋巴道转移中有着重要意义,HLA-B表达可作为一项监测OSCC恶性程度与淋巴道转移的指标。     

Expressions of CXCR7/ligands may be involved in oral carcinogenesis

Oral carcinogenesis is a multistep process and requires accumulation and interplay of a series of molecular events. Chemokines and their receptors have been suggested to play important roles in the initiation or progression of cancers. Until now, no report focuses on their alterations in premalignant stage of oral squamous cell carcinoma (OSCC). Compared with normal tissues, mRNA levels of 9 chemokines and 3 chemokine receptors including CXCR7 in oral leukoplakia (OLK) were increased more than two folds by microarray analysis. Then, CXCR7 was selected for further confirmation and immunohistochemistry examination during multistage oral carcinogenesis. CXCR7 was expressed in 85% of OLK and 86% of OSCC. However, only 8% (1 of 13 cases) of normal tissue displayed CXCR7 immunostaining. The positive ratios of CXCR7, CXCL12 and CXCL11 in OLK and OSCC tissues respectively, were significantly higher than that in normal epithelia (P < 0.05), although no significant difference was found between OLK and OSCC. Meanwhile, CXCR7 always concomitantly expressed with it ligands in OLK and OSCC tissues. Our results indicated that CXCR7-CXCL12/CXCL11 axis might play important roles in oral carcinogenesis.     

Quantitative Risk Stratification of Oral Leukoplakia with Exfoliative Cytology

Exfoliative cytology has been widely used for early diagnosis of oral squamous cell carcinoma (OSCC). Test outcome is reported as “negative”, “atypical” (defined as abnormal epithelial changes of uncertain diagnostic significance), and “positive” (defined as definitive cellular evidence of epithelial dysplasia or carcinoma). The major challenge is how to properly manage the “atypical” patients in order to diagnose OSCC early and prevent OSCC. In this study, we collected exfoliative cytology data, histopathology data, and clinical data of normal subjects (n=102), oral leukoplakia (OLK) patients (n=82), and OSCC patients (n=93), and developed a data analysis procedure for quantitative risk stratification of OLK patients. This procedure involving a step called expert-guided data transformation and reconstruction (EdTAR) which allows automatic data processing and reconstruction and reveals informative signals for subsequent risk stratification. Modern machine learning techniques were utilized to build statistical prediction models on the reconstructed data. Among the several models tested using resampling methods for parameter pruning and performance evaluation, Support Vector Machine (SVM) was found to be optimal with a high sensitivity (median>0.98) and specificity (median>0.99). With the SVM model, we constructed an oral cancer risk index (OCRI) which may potentially guide clinical follow-up of OLK patients. One OLK patient with an initial OCRI of 0.88 developed OSCC after 40 months of follow-up. In conclusion, we have developed a statistical method for qualitative risk stratification of OLK patients. This method may potentially improve cost-effectiveness of clinical follow-up of OLK patients, and help design clinical chemoprevention trial for high-risk populations.     

Comprehensive bioinformatics analysis combined with experimental validation to screen biomarkers for malignant transformation of oral leukoplakia

Oral leukoplakia (OLK) is the most common potentially malignant disorders in the oral cavity. This study aimed to screen the key genes of OLK malignant transformation using the Gene Expression Omnibus (GEO) database and experiments. In this study, the GEO database was employed to screen OLK malignant transformation-related genes, which were subsequently identified with a series of bioinformatic analyses. External validation showed that the model based on LAPTM4B, NR3C1, and COX6A1 had high accuracy in diagnosing OLK malignant transformation. Furthermore, the DMBA-induced potentially malignant disorders and OSCC models in vivo and real-time PCR experiment in vitro further verified the database analysis results. In conclusion, three key genes (LAPTM4B, NR3C1, and COX6A1) were screened as potential biomarkers for the diagnosis and treatment of OLK malignant transformation.     

Urine metabolite profiling offers potential early diagnosis of oral cancer

Oral cancer is the sixth most common human cancer, with a high morbidity rate and an overall 5-year survival rate of less than 50%. It is often not diagnosed until it has reached an advanced stage. Therefore, an early diagnostic and stratification strategy is of great importance for oral cancer. In the current study, urine samples of patients with oral squamous cell carcinoma (OSCC, n = 37), oral leukoplakia (OLK, n = 32) and healthy subjects (n = 34) were analyzed by gas chromatography-mass spectrometry (GC–MS). Using multivariate statistical analysis, the urinary metabolite profiles of OSCC, OLK and healthy control samples can be clearly discriminated and a panel of differentially expressed metabolites was obtained. Metabolites, valine and 6-hydroxynicotic acid, in combination yielded an accuracy of 98.9%, sensitivity of 94.4%, specificity of 91.4%, and positive predictive value of 91.9% in distinguishing OSCC from the controls. The combination of three differential metabolites, 6-hydroxynicotic acid, cysteine, and tyrosine, was able to discriminate between OSCC and OLK with an accuracy of 92.7%, sensitivity of 85.0%, specificity of 89.7%, and positive predictive value of 91.9%. This study demonstrated that the metabolite markers derived from this urinary metabolite profiling approach may hold promise as a diagnostic tool for early stage OSCC and its differentiation from other oral conditions.     

The enigmatic role of matrix metalloproteinases in epithelial-to-mesenchymal transition of oral squamous cell carcinoma: Implications and nutraceutical aspects

The most prevalent malignancy in the oral cavity is represented by oral squamous cell carcinoma, an aggressive disease mostly detected in low-income communities. This neoplasia is mostly diffused in older men particularly exposed to risk factors such as tobacco, alcohol, and a diet rich in fatty foods and poor in vegetables. In oral squamous cell carcinoma, a wide range of matrix-cleaving proteinases are involved in extracellular matrix remodeling of cancer microenvironment. In particular, matrix metalloproteinases (MMPs) represent the major and most investigated protagonists. Owing to their strong involvement in malignant pathologies, MMPs are considered the most promising new biomarkers in cancer diagnosis and prognosis. The interest in studying MMPs in oral cancer biology is also owing to their prominent role in epithelial-to-mesenchymal transition (EMT). EMT is an intricate process involving different complex pathways. EMT-related proteins are attractive diagnostic biomarkers that characterize the activation of biological events that promote cancer's aggressive expansion. Different antioncogenic natural compounds have been investigated to counteract oral carcinogenesis, with the scope of obtaining better clinical results and lower morbidity. In particular, we describe the role of different nutraceuticals used for the regulation of MMP-related invasion and proliferation of oral cancer cells.     

口服轮状病毒活疫苗的安全性临床观察

为了观察口服轮状病毒活疫苗的安全性,2004年4~10月在济南市中心医院预防接种门诊对总服用该疫苗的410例7月龄~3岁的幼儿密切观察其反应,测量腋下体温并询问有无不适症状等。结果显示,95.61%幼儿(392/410)无不良反应;约4.39%(18/410)出现发热、腹泻等不良副反应,其中强反应3例(0.73%),中度反应11例(2.68%),轻度反应4例(0.98%),均于对症治疗后3日内缓解并消失。临床资料证明,使用该疫苗是安全的。     

Gene Expression in Tilapia Following Oral Delivery of Chitosan-Encapsulated Plasmid DNA Incorporated into Fish Feeds

DNA delivery into fish is important for transient gene expression, (e.g., DNA vaccination). Previous studies have generally focused on intramuscular injection of DNA vaccines into fish. However, this method is obviously impractical and laborious for injecting large numbers of fishes. This study reports oral delivery of a construct expressing the β-galactosidase reporter gene into fish by encapsulating the DNA in chitosan and incorporating it into fish feeds. We found that β-galactosidase expression could be observed in the stomachs, spleens, and gills of fishes fed with flakes containing the chitosan-DNA complex. These results suggest that DNA vaccines and other constructs can be easily and cheaply delivered into fishes orally by use of carriers and incorporation into fish feeds.