口腔癌细胞诱导的淋巴管内皮细胞基因表达谱的变化

目的探讨在口腔癌细胞影响下淋巴管内皮细胞分子表型的变化。方法利用体外共培养模型,将淋巴管内皮细胞与口腔癌细胞进行共培养,模拟肿瘤中淋巴管内皮细胞的变化。应用Affymetrix U133 Plus 2.0基因芯片对肿瘤组织中淋巴管内皮细胞（TLEC）与淋巴管内皮细胞（LEC）基因表达的差异进行了检测和比较。并对功能相似的表达差异基因进行分类。结果差异基因表达谱共发现了677个基因表达差异在1倍以上,其中在TLEC中表达上调的基因有384条,下调的基因有293条。这些基因与细胞黏附、凋亡、运动、发育及血管生成有关。同时这些基因还参与细胞的信号传导、免疫应答、细胞代谢等过程。结论LEC与TLEC在分子水平是有差别的。以此为基础,可以针对淋巴管内皮细胞进行靶向阻断,达到治疗口腔癌淋巴道转移的目的。     

p53在口腔癌癌变过程中的研究

肿瘤抑制基因 p5 3的突变是口腔癌中最常见的基因改变之一。研究 p5 3基因与蛋白在口腔癌变过程中的改变 ,有助于口腔癌的早期诊断、预防 ,并对其预后及治疗效果的判定有指导意义。     

口腔扁平苔藓癌变的临床分析

作者对经临床和组织学确诊的６３３例口腔扁平苔藓患者进行１～１７年随访，有８例扁平苔藓癌变，癌变率为１．３％。其中男性３例，占男性扁平苔藓数的１．６％；女性５例，占女性扁平苔藓数的１．１％。癌变组平均年龄为５０．９岁，病史平均为１０．９年；癌变损害部位分别为颊部、下唇和舌腹缘；损害类型均为糜烂型，１例伴斑块增殖。癌变组术后随访到７例，平均随访３．６年，有２例癌肿复发。     

用比较蛋白质组学技术筛选口腔黏膜上皮细胞体外癌变的相关蛋白

目的：探讨口腔黏膜上皮细胞在体外癌变不同阶段表达的差异蛋白质。方法：以口腔黏膜上皮细胞体外癌变模型为对象,采用双向凝胶电泳技术和图像分析软件PDQuest分离和分析不同阶段细胞间的差异蛋白质点,采用LC-MS/MS质谱分析系统鉴定差异蛋白质点,采用Gene Ontology Annotation将已知差异蛋白质进行分类。结果：采用双向凝胶电泳技术和图像分析软件PDQuest,得到差异蛋白质点54个,采用LC-MS/MS质谱鉴定后,共得到候选差异蛋白质45个。根据Gene Ontology Annotation分类,按细胞组成分布最多的差异蛋白质位于细胞质和细胞膜,按分子功能分布最多的是磷酸酶活性、催化活性、结构分子功能和钙离子结合功能,按生物过程分布最多的是代谢过程、细胞信号传导和细胞黏附与运动。结论：比较蛋白质组学方法中的双向凝胶电泳和质谱技术,能够很好分离和鉴定口腔黏膜上皮细胞体外癌变模型中不同阶段细胞的差异蛋白质。     

苯丙芘联合佛波酯恶性转化HIOEC细胞的实验研究

目的:诱导永生化口腔上皮细胞系HIOEC细胞恶性转化,建立HIOEC-B(a)P-TPA细胞系。方法:通过递增浓度的化学致癌剂苯丙芘复合促癌剂佛波酯体外诱导HIOEC细胞,筛选得到具有恶性表型的鳞状细胞癌细胞系HIOEC-B(a)P-TPA。通过微分干涉显微镜和HE染色观察细胞形态学改变,免疫组化染色检测细胞角蛋白和波形蛋白表达情况;软琼脂集落形成、裸小鼠异体皮下成瘤性鉴定HIOEC-B(a)P-TPA细胞的恶性表型。结果:(1)HIOEC细胞诱导培养6个月后,可以在正常生理钙离子浓度、含胎牛血清培养液中生长;(2)细胞在诱导过程中形态多变,最后以成纤维样细胞为主,异形性明显;(3)HIOEC-B(a)P-TPA细胞中细胞角蛋白表达减弱,同时表达波形蛋白;(4)有很强的软琼脂集落形成能力,集落形成率为24.5%;(5)HIOEC-B(a)P-TPA细胞尚未观察到成瘤性。结论:生物化学致癌因素B(a)P和TPA可诱导HIOEC细胞恶性转化,反映了口腔黏膜上皮细胞体外恶变发生、发展过程。     

口腔扁平苔藓基因表达谱中差异表达基因初步探讨

目的筛选口腔扁平苔藓组织中的差异表达基因,并对其进行功能分类。方法用4 000种人类基因多聚酶链反应产物制成BiostarH- 40s型表达谱芯片,分离纯化正常口腔黏膜组织和口腔扁平苔藓病变组织mRNA,制备表达谱探针,用ScanArray 4000 荧光扫描仪扫描芯片荧光信号图像,分析正常口腔黏膜组织和口腔扁平苔藓组织之间差异表达的基因。结果1）在4 000条基因中,有213条基因表达差异,其中122条基因表达上调,91条基因表达下调。2）在表达上调基因中,功能分类主要包括免疫相关基因、代谢相关基因、癌基因、细胞因子、细胞信号和传递蛋白。3）在表达下调基因中,功能分类主要包括DNA结合、转录和转录因子、细胞信号和传递蛋白、免疫相关基因、细胞因子、代谢相关基因。结论口腔扁平苔藓的发生、发展过程中存在着多条不同功能基因表达调控的改变。     

口腔黏膜癌变过程中Fas与FasL表达的相互关系

目的　初步探讨Fas和Fas L蛋白在口腔黏膜癌前损害发生发展过程中的作用及变化规律。方法　分别选出正常口腔黏膜、上皮单纯增生、上皮轻度异常增生、上皮中度异常增生、上皮重度异常增生与原位癌、鳞状细胞癌标本,共64例,采用免疫组织化学染色技术---酶标链亲和素生物素法(LsAB)染色并进行光镜下观察。结果　多数口腔鳞癌显示Fas表达的下调和Fas L表达的上调,同样的结果见于口腔癌前损害黏膜组织。结论　Fas 和Fas L的表达参与了口腔癌变过程,且可能是癌细胞逃避宿主免疫攻击的机制。Fas/Fas L系统有望成为判定口腔癌前损害预后的生物标志。     

口腔黏膜癌变过程中血清生化标志物的研究进展

口腔黏膜癌变是指口腔黏膜从正常状态经口腔潜在恶性疾患阶段发展为口腔鳞状细胞癌,直至远处转移的多步骤、多阶段、多基因改变的过程。该过程涉及多种血清生化标志物的变化,这些动态变化对于病情的进展具有重要的提示作用,对临床有着深远的意义。本文将与口腔黏膜癌变过程相关的血清生物标志物的研究现状作一综述。     

口腔扁平苔藓组织中p73和p14^ARF基因表达与癌变的关系

目的探讨p73、p14^ARF在口腔扁平苔藓组织中的表达与癌变的关系。方法采用免疫组化方法,分别检测口腔扁平苔藓、口腔鳞状细胞癌和正常口腔粘膜各30例组织中p73、p14^ARF的表达情况;采用美国Media Cybernetics公司的Image Plus5.0专业图像分析软件进行IOD值（累计光密度值）的测量。结果p73表达的IOD值在正常口腔粘膜、口腔扁平苔藓及口腔鳞状细胞癌中不断增高,而p14ARF表达的IOD值在正常口腔粘膜、口腔扁平苔藓及口腔鳞状细胞癌中却逐渐下降。两组间有统计学意义（P〈0.05）。p73、p14^ARF在口腔扁平苔藓、正常口腔粘膜、口腔鳞状细胞癌组织中的表达呈负性相关性（P〈0.05）。结论p73、p14^ARF这两个基因在OLP的癌变过程中具有一定的作用。     

Characteristics of a cancerous cell line, HIOEC-B(a)P-96, induced by benzo(a)pyrene from human immortalized oral epithelial cell line

Oral squamous cell carcinoma (OSCC) is the most common malignant tumor in the oral and maxillofacial region. The mechanism of carcinogenesis of OSCC is still unclear. In vitro study on OSCC cell lines, especially derived from immortalized oral epithelial cells, is a very useful strategy to understand the mechanism of carcinogenesis. Based on our previous human immortalized oral epithelial cell (HIOEC) line, obtained from normal oral epithelial cells by transfection of HPV16 E6/E7 gene, a new cancerous cell line, HIOEC-B(a)P-96 (HB96), was established from the HIOEC by induction with benzo(a)pyrene. The characteristics of the HB96 cells such as cell morphology, ultrastructure, proliferation ability, invasion ability, and tumorigenesis were studied. The HB96 cells lost contact inhibition with uncontrolled cell division and obvious cell overlap, they were polygonal in shape and ununiform in size with increased ratio between nucleus and plasma. Increased proliferative ability and invasion ability were confirmed by the cell proliferation analysis and cell invasion assay, respectively. The tumorigenicity of well to moderately differentiated squamous cell carcinoma was confirmed in the nude mice experiments pathologically. Increased expression of HPV16 E6/E7 proteins and obvious correlation with decreased expression of p53 and Rb proteins was also confirmed by Western blotting. Thus, this HB96 cell line induced by benzo(a)pyrene from the HIOEC line is a useful tool to study the mechanism of carcinogenesis of OSCC in vitro for future genomic and proteomic analyses. It is also the first in vitro cancerous cell line of OSCC in China derived from immortalized oral epithelial cells.     

口腔黏膜上皮细胞体外癌变模型的建立

目的诱导永生化口腔上皮细胞系（HIOEC）细胞恶性转化，建立口腔黏膜上皮细胞体外癌变模型。方法通过化学致癌剂苯并芘[B（a）P]体外诱导HIOEC细胞，逐步筛选到鳞状细胞癌细胞系HIOEC-B（a）P。通过微分干涉显微镜和HE染色观察细胞形态学改变；用软琼脂集落形成及裸小鼠异体皮下成瘤的实验鉴定HIOEC-B（a）P细胞的恶性表型。结果①HIOEC细胞在B（a）P诱导培养后可以在正常生理钙离子浓度、含胎牛血清培养液中生长；②细胞在诱导过程中形态多变，最后稳定为多角形铺路石样上皮细胞；③第93代HIOEC-B（a）P细胞开始具备软琼脂集落形成能力；④HIOEC—B（a）P第55代细胞在裸小鼠皮下首次形成角化团块，第69代细胞形成分化较好的典型鳞状细胞癌，第74代、第96代细胞均形成与临床病理相似的Ⅰ～Ⅱ级鳞状细胞癌。结论生物因素合并化学致癌因素B（a）P可以诱导永生化口腔黏膜上皮细胞恶性转化，为进一步研究口腔黏膜上皮细胞多因素、多步骤癌变机制提供实验模型。     

TNFAIP6在口腔疣状癌与口腔鳞状细胞癌中的表达及其与临床病理的关系

目的： 研究口腔疣状癌（OVC）及口腔鳞状细胞癌（OSCC）中肿瘤坏死因子α诱导蛋白6（TNFAIP6）的表达其与临床病理的关系。方法： 在TCGA数据库中下载OSCC的RNA-seq数据,采用两样本t检验比较TNFAIP6在OSCC和正常口腔黏膜中的表达差异,采用受试者工作曲线（ROC）和生存曲线分别评价其诊断和预后价值。取15例正常口腔黏膜、22例OVC和24例OSCC及对应癌旁组织,利用免疫组织化学方法检测上述标本中TNFAIP6的表达和分布,并分析其阳性表达率与临床病理因素的关系。取5例OSCC、3例OVC及对应癌旁组织,利用反转录聚合酶链反应、免疫印迹技术验证其转录及蛋白表达水平,使用Graph Pad Prism 7.0软件进行计量数据分析,采用SPSS 21.0软件包对各组间数据进行两样本均数t检验。结果： 在TCGA数据库中,TNFAIP6在OSCC中表达显著上调且具有一定的诊断价值,然而其表达失调与OSCC患者总体生存率无关。在OSCC石蜡标本中,TNFAIP6表达显著上调,与饮酒、颈淋巴结转移、临床分期有关。在OVC石蜡标本中,TNFAIP6表达显著上调;TNFAIP6在OSCC癌组织的转录及蛋白表达显著上调。结论： TNFAIP6在OSCC组织中异常高表达,并与诊断及预后相关,可作为肿瘤分子标志物。     

Chemopreventive activity of apple extract following medium-term oral carcinogenesis assay induced by 4-nitroquinoline-1-oxide

ObjectiveThe aim of this study was to evaluate the chemopreventive activity of an apple extract following medium-term oral carcinogenesis assay induced by 4-nitroquinoline-1-oxide (4NQO).MethodsA total of 30 male Wistar rats were distributed into five groups as follows (n = 6 per group): Group 1, negative control group (non-treated group); Group 2, received 4NQO during 8 weeks in drinking water and treated with apple extract at 1% by gavage between the first and fourth weeks daily (initiation phase); Group 3, received 4NQO for 8 weeks in drinking water and treated with apple extract by gavage at 1% between the fifth and eighth weeks daily (promotion phase); Group 4, received apple extract at 1% by gavage for 8 consecutive weeks only; and Group 5, received 4NQO for 8 weeks in drinking water daily.ResultsHistopathological analysis revealed decreased hyperplasic lesions in Group 2 when compared with Group 5. Likewise, decreased dysplastic lesions in Group 3 were observed when compared with Group 5. In Groups 2 and 3, decreased COX-2 and TNF-alpha gene expressions were observed when compared with Group 5. Cytochrome c and caspase 3 levels increased in Groups 2 and 3 when compared with Group 5.ConclusionIn conclusion, our results demonstrate that apple extract suppresses rat tongue carcinogenesis as a result of anti-inflammatory activity and apoptosis through the intrinsic mitochondrial pathway.     

Dental attendance of oral and oropharyngeal cancer patients in a public hospital in Western Australia

Background: Dentists have recently seen the introduction of devices which aim to facilitate early oral cancer detection, sparking interest in opportunistic oral cancer screening. However, concerns have been raised about the lack of regular dental attendance amongst high risk individuals. The purpose of this study was to obtain information pertaining to dental attendance of oral and oropharyngeal cancer patients. Methods: All records of patients referred to the Oral Medicine Clinic at the Oral Health Centre of Western Australia, between January 2005 and December 2009, from one major teaching hospital were examined. Information extracted included age, gender, smoking status, referral date, tumour type, tumour site, disease stage (TNM classification), and information on dental attendance. Outcomes measured included time (months) since the patient’s last dental visit and information concerning regularity of dental attendance. Results: No association was found between dental attendance and gender, smoking, disease stage or age at diagnosis. Most patients had not visited a dentist in the preceding 12 months. The mean date of last dental visit was 5.6 years prior. Conclusions: More should be done in Australia to encourage patients at high risk of oral cancer to attend the dentist and undergo annual oral soft tissue examination.     

前列腺素E2和白三烯B4对口腔癌促进作用的研究

目的通过在DMBA诱导的金黄地鼠口腔癌颊囊模型上,局部应用白三烯n4（LTB4）或前列腺素E2（PGE2）,探讨LTB4和PGE2在口腔癌形成中的作用,进一步阐明口腔癌的发病机制。方法用DMBA涂抹金黄地鼠左侧颊囊粘膜4周,第五周开始分别局部涂抹LTB4和PGE220周,处死动物,取全部颊囊进行肉眼观察计数和组织学检查,并进行统计学分析。结果LTB4＋DMBA组癌变率为51.4％（18／35）;PGE2＋DMBA组癌变率为45.7％（16／35）,均显著高于DMBA组20.0％（7／35）。另外两组不使用DMBA启动,同时局部分别涂抹LTB4和PGE220周,两组均有异常增生发生,但是没有癌变。结论PGE2和LTB4是口腔癌形成的促进剂,并且有可能成为化学预防作用的重要靶点。     

组织蛋白酶B在口腔黏膜上皮癌变过程中的表达及意义

目的：探讨组织蛋白酶B（Cathepsin B,CB）在口腔鳞状细胞癌中的表达及临床意义。方法：以口腔黏膜上皮永生化细胞系（human immortalized oral epithelia cell line,HIOEC）及经过苯丙芘[benzo（a）pyrene,B（a）P]诱导产生鳞状细胞癌细胞（HB）而形成的口腔鳞癌体外癌变模型为研究对象,通过双向凝胶电泳（2-DE）和质谱分析（LC-MS/MS）筛选和鉴定出差异蛋白质CB。采用实时定量PCR、Western印迹和免疫组化方法,检测口腔鳞癌细胞和30例原发口腔鳞癌标本中的mRNA及蛋白质表达水平。采用SPSS10.0软件包对数据进行非参数检验。结果：与永生化HIOEC相比,HB和CAL27细胞中CB的mRNA水平显著升高,HB、Tca8113、TSCC、CAL27和OSC细胞中CB蛋白表达水平明显升高。30例口腔鳞癌患者癌组织中CB的mRNA和蛋白表达水平均较癌旁组织升高（P〈0.01）。CB的表达水平与肿瘤大小、淋巴结转移、临床分期、病理分化程度无显著相关性。结论：CB在口腔鳞癌中的表达明显升高,提示其与肿瘤的发生、发展具有一定关系。     

Increase in detection of oral cancer and precursor lesions by dentists: Evidence from an oral and maxillofacial pathology service

BackgroundDentists play an important role in the detection and diagnosis of oral diseases, including oral cancer and its precursor lesions. There are few comprehensive reviews in the recent literature that examine the scope and trends of oral disease diagnoses by dentists.MethodsThe authors analyzed all accessions to the Toronto Oral Pathology Service at the Faculty of Dentistry at the University of Toronto in Toronto, Ontario, Canada, from 2005 through 2015 using a custom-built database. They used these data to calculate the temporal trends in the diagnoses of oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC).ResultsA total of 63,483 biopsy specimens were submitted primarily by dentists from 2005 through 2015. From these, 2,679 cases of OED and 828 OSCC were diagnosed. The authors’ results show a 3.8-fold increase in the number of epithelial dysplasias and a 1.8-fold increase in mucosal carcinomas over the study period. The rate of increase of OED and OSCC was significantly higher than the rate of increase of total oral carcinomas diagnosed in the region, the population changes, and the number of dentists in the region.Conclusions and Practical ImplicationsWithin the limitations of a study of a single large oral pathology biopsy service, the analysis of diagnoses shows that dentists are increasingly involved in the detection of oral mucosal carcinoma and precursor lesions. The dental community plays an important and increasing role in the detection of oral cancer and potentially malignant disorders. Increased awareness among oral health care and nonoral health care professionals may increase early detection of OSCC.