Patients with oral cancer developing from pre‐existing oral leukoplakia: do they do better than those with de novo oral cancer?

Background:  It has been suggested that patients with squamous cell carcinomas derived from oral leukoplakia have a better prognosis than patients with carcinomas that are not associated with oral leukoplakia.
Aim:  To study the mortality rate of 19 patients with a squamous cell carcinoma derived from pre-existing oral leukoplakia.
Method:  The mortality rate of 19 patients with a proven oral squamous cell carcinoma derived from a pre-existing oral leukoplakia was compared with that of a similar size group of patients with oral carcinoma without a pre-existing oral leukoplakia, being matched for gender, age, smoking habits, use of alcohol, oral subsite and histopathologic grade. Treatment in all patients was primarily by surgical excision. The mortality rates up to 5 years have been computed according to the Kaplan–Meier method.
Result:  No significant difference of the mortality rates up to 5 years of follow-up was observed between the two groups of patients.
Conclusion:  Patients with oral cancer developing from pre-existing oral leukoplakia do not do better than those with de novo oral cancer.     

Human papillomavirus infection in oral potentially malignant disorders and cancer

Human papillomavirus (HPV) infects keratinocytes in the mucosa or skin, and persistent infection with HPV may lead to premalignant lesions and invasive cancer, especially cervical cancer. It has also been hypothesized that HPV infection is an etiological factor of oral squamous cell carcinoma and oral precancerous disorders such as lichen planus, leukoplakia, and erythroplakia. A high percentage of HPV in oral lesions supports the possible viral contribution, but an association of HPV infection with these lesions remains to be established. The current paper will update the latest progress of HPV infection in several oral potentially malignant disorders and oral squamous cell carcinoma and discuss the impact of HPV infection on the progression of oral potentially malignant disorders.     

Mutated and wild-type p53 expression and HPV integration in proliferative verrucous leukoplakia and oral squamous cell carcinoma

The frequencies of overexpression and mutation in the p53 tumor suppressor gene were examined in proliferative verrucous leukoplakia and oral squamous cell carcinoma with immunohistochemistry and single-strand conformation polymorphism analysis of DNA fragments amplified by polymerase chain reaction. Ten samples each of normal oral mucosa, proliferative verrucous leukoplakia, and squamous cell carcinoma were immunostained with antibodies against p53 protein; 8 of 10 cases of proliferative verrucous leukoplakia cases and 7 of 10 cases of oral squamous cell carcinoma were positive for p53 protein. Minimal staining was observed in normal oral tissues. The quantified labeling indexes demonstrated a range that corresponded to lesion progression. Single-strand conformation polymorphism analysis revealed p53 gene mutations within exons 5 to 8 in 40% (4 of 10) of the squamous cell carcinoma samples. Two of the 4 mutated squamous cell carcinoma samples lacked p53 expression. No p53 mutations were detected in proliferative verrucous leukoplakia tissues. Human papillomavirus 16 was identified in 2 of 7 p53 positive oral squamous cell carcinoma samples. Human papillomavirus 16 and 18 were identified in two of eight p53 positive proliferative verrucous leukoplakia samples. One p53 negative squamous cell carcinoma sample was positive for human papillomavirus 16 and had a mutation in exon 6 of the p53 gene. Human papillomavirus infection along with p53 expression plays a yet to be defined role in the pathogenesis of a limited number of cases of proliferative verrucous leukoplakia and squamous cell carcinoma. p53 Immunohistochemistry, p53 gene mutations, and human papillomavirus infection prevalence do not provide a means to differentiate between leukoplakia and carcinoma and do not provide a predictive test for progression of leukoplakia to carcinoma.     

Human papillomavirus as a risk factor for oral squamous cell carcinoma: a meta-analysis, 1982-1997

OBJECTIVE: Human papillomavirus (HPV) infection is a significant risk factor for uterine cervical carcinoma. However, the role of HPV infection in oral squamous cell carcinoma (OSCC) is less well defined. To determine the significance of the relationship of this virus in the progressive development of oral cancer, we estimated the risk of HPV detection in normal oral mucosa, precancerous oral tissue, and oral carcinoma using meta-analysis. STUDY DESIGN: Case reports and clinical series published in English-language journals were retrieved by searching MEDLINE (January 1980-August 1998). Review articles were also examined to identify additional studies. Studies that used biochemical, immunologic, microscopic, or molecular analyses to detect HPV in tissue or cells derived from normal oral mucosa (n = 25), benign leukoplakia (n = 21), intraepithelial neoplasia (ie, dysplasia and carcinoma in situ; n = 27), and oral cancer (n = 94) were included in the meta-analysis. Information on sample size, age, sex, method of tissue preservation (ie, fresh, frozen, paraffin-embedded), assay, primer amplification region (early, late), high-risk versus low-risk genotype, and use of tobacco or alcohol was abstracted by one author (C.S.M.). RESULTS: Data from 94 reports that analyzed 4680 samples were included in the meta-analysis. Analyses made by means of a random-effects model with and without adjustments for assay sensitivity showed increased probability of HPV detection in tissue with precancerous and cancerous features compared with normal mucosa. The likelihood of detecting HPV in normal oral mucosa (10.0%; 95% confidence interval [CI], 6.1%-14.6%) was significantly less than of detecting benign leukoplakia (22.2%; 95% CI, 15.7%-29.9%), intraepithelial neoplasia (26.2%; 95% CI, 19.6%-33.6%), verrucous carcinoma (29.5%; 95% CI, 23%-36.8%), and OSCC (46.5%; 95% CI, 37.6%-55.5%). Adjustment of findings for differences in assay sensitivity indicated that these estimates may be conservative. Overall, HPV was between 2 and 3 times more likely to be detected in precancerous oral mucosa and 4.7 times more likely to be detected in oral carcinoma than in normal mucosa. The pooled odds ratio for the subset of studies directly comparing the prevalence of HPV in normal mucosa and OSCC was 5.37, confirming the trend observed in the overall sample. The probability of detecting high-risk HPVs in OSCCs was 2.8 times greater than that of low-risk HPVs. CONCLUSION: This meta-analysis indicates that HPV is detected with increased frequency in oral dysplastic and carcinomatous epithelium in comparison with normal oral mucosa. The findings provide further quantitative evidence that oral infection with HPV, particularly with high-risk genotypes, is a significant independent risk factor for OSCC.     

Short telomeres in an oral precancerous lesion: Q-FISH analysis of leukoplakia

J Oral Pathol Med (2012) 41 : 372–378 Objectives: A precancerous condition is a lesion that, if left untreated, leads to cancer or can be induced to become malignant. In the oral region, leukoplakia is a lesion that has been regarded as precancerous. In cases of oral carcinoma, we have frequently noticed that a type of leukoplakia histologically demonstrating hyper‐orthokeratosis and mild atypia (ortho‐keratotic dysplasia; OKD) is often associated with carcinoma, either synchronously or metachronously. Therefore, we consider OKD‐type leukoplakia to be a true precancerous lesion. Materials and Methods: In an attempt to clarify the relationship between OKD as a precancerous condition in the oral mucosa and telomere length, we estimated telomere lengths in this type of leukoplakia using quantitative fluorescence in situ hybridization, and also quantified the frequency of anaphase–telophase bridges (ATBs) in comparison with squamous cell carcinoma in situ (CIS) and the background tissues of CIS and OKD. Results: Ortho‐keratotic dysplasia was frequently associated with squamous cell carcinoma (45.0%) and showed significantly shorter telomeres than normal control epithelium, CIS, or the background of CIS or OKD. The frequency of ATBs was much higher in OKD than in control epithelium or CIS. Conclusion: Ortho‐keratotic dysplasia appears to be frequently associated with carcinoma, chromosomal instability, and excessively shortened telomeres, not only in the lesion itself but also in the surrounding background. Therefore, when this type of leukoplakia is recognized in the oral region, strict follow‐up for oral squamous cell carcinoma is necessary, focusing not only on the areas of leukoplakia, but also the surrounding background.     

CC趋化因子7和趋化因子受体7在口腔白斑及口腔鳞癌组织中的表达及意义

目的 :探讨CC趋化因子7在口腔白斑组织中的表达及意义。方法 :选取2011-03—2014-03期间,在我院口腔科治疗的口腔白斑患者43例,口腔鳞癌患者41例,以及正常口腔黏膜组织46份,利用免疫组化法对不同组织中CCL7和CCR7蛋白表达情况进行检测。结果 :CCL7蛋白在口腔白斑组织中的表达低于口腔鳞癌组织,而高于正常黏膜组织,差异有统计学意义(P<0.05);CCR7蛋白在口腔白斑组织中表达低于口腔鳞癌组织,而高于和正常黏膜组织,差异有统计学意义(P<0.05);Spearman相关分析显示,口腔白斑组织中CCL7蛋白和CCR7蛋白表达呈正相关(r=0.594,P<0.05)。结论:CCL7和CCR7蛋白表达均表现出从口腔正常黏膜组织到口腔白斑到口腔鳞癌逐渐升高的趋势,且二者呈正相关,可能参与了口腔白斑癌变过程。     

原位杂交、免疫组化及AgNOR染色法在口腔念珠菌性白斑诊断中的应用

目的:探讨原位杂交、免疫组化及AgNOR染色法在口腔念珠菌性白斑(OCL)诊断中的综合应用。方法:采用原位杂交、免疫组化及AgNOR染色法,并对照PAS染色法,检测154例口腔白斑及鳞癌组织中念珠菌的感染情况。结果:经统计学处理,4种检测方法之间无显著性差异;白斑真菌感染率为71.8%(89/124),白色念珠菌感染率为56.5%(70/124)。本研究中只发现1例热带念珠菌感染。真菌感染中78.7%(70/89)为白色念珠菌。鳞癌真菌感染率63.3%(19/30),均为白色念珠菌感染。结论:AgNOR染色法、原位杂交和免疫组化的综合应用,可以从组织、细胞、分子水平对OCL进行定位研究,为OCL的准确诊断提供客观的实验依据。     

Nested PCR for detection of HSV-1 in oral mucosa

Background

It has been estimated that 15%-20% of human tumours are driven by infection and inflammation, and viral infections play an important role in malignant transformation. The evidence that herpes simplex virus type 1 (HSV-1) could be involved in the aetiology of oral cancer varies from weak to persuasive. This study aimed to investigate by nested PCR (NPCR) the prevalence of HSV-1 in samples from normal oral mucosa, oral leukoplakia, and oral squamous cell carcinoma (OSCC).

Material and Methods

We investigated the prevalence of HSV-1 in biopsies obtained from 26 fresh, normal oral mucosa from healthy volunteers as well as 53 oral leukoplakia and 27 OSCC paraffin-embedded samples. DNA was extracted from the specimens and investigated for the presence of HSV-1 by nested polymerase chain reaction (NPCR) and DNA sequencing.

Results

HSV-1 was detected in 14 (54%) of the healthy samples, in 19 (36%) of the oral leukoplakia samples, and in 14 (52%) of the OSCC samples. The differences were not statistically significant.

Conclusions

We observed a high incidence of HSV-1 in healthy oral mucosa, oral leukoplakia, and OSCC tissues. Thus, no connection between OSCC development and presence of HSV-1 was detected. Key words:HSV-1, nested PCR, PCR.     

表面增强激光解吸-电离质谱法筛选唾液中口腔鳞状细胞癌蛋白标志物

目的 在唾液中筛选口腔癌前病变、鳞状细胞癌、转移癌并与健康人鉴别的肿瘤蛋白标志物.方法 在CM-10蛋白质芯片上采用表面增强激光解吸-电离(Surface enhanced laser desorption/ionization(SELDI)质谱法技术对口腔白斑(6例)、鳞状细胞癌(17例)、转移癌(7例)和健康者(15人)的非刺激性全唾液中的蛋白标志物进行检测,支持向量机法建立指纹图谱诊断模式.结果 口腔鳞状细胞癌和健康人唾液蛋白鉴别模式:蛋白质荷比峰(简称:质荷比)5797、2902、3883和4951组合,敏感性为88.24%、特异性为93.33%;口腔鳞状细胞癌和白斑鉴别模式:质荷比为5818、4617和3884的组合,敏感性为100.00%、特异性为100.00%;口腔鳞状细胞癌和局部转移癌的鉴别模式:质荷比为55 809和5383的组合,敏感性为94.12%、特异性为85.71%.结论 通过SELDI质谱法技术筛选出的肿瘤标志物可以辅助口腔鳞状细胞癌的早期诊断,预测白斑向鳞状细胞癌转化及癌局部转移的潜能.     

不同癌基因在口腔白斑和鳞癌中表达的研究

目的 研究正常口腔黏膜，口腔黏膜白斑及口腔鳞癌组织中不同癌基因的表达。方法 通过HE染色确定口腔黏膜组织类型，提取组织中RNA，采用RT－PCR的方法研究不同组织中c-myc、ras、cyclinD1及bcl-2基因的表达。结果 正常口腔黏膜中未见c-myc、ras、cyclinD1及bcl-2基因表达；轻、中、重度异常增生白斑和鳞癌组织中c-myc、ras、cyclinD1及bcl-2基因表达显著增加。结论 c-myc、ras、cyclinD1及bcl-2基因的表达增加与口腔黏膜白斑及鳞癌的发生相关。     

TRAIL在口腔鳞状细胞癌和白斑中的表达及意义

目的：研究肿瘤坏死因子相关凋亡诱导配体（TNF-related apoptosis inducing ligand,TRAIL）在口腔鳞状细胞癌（OSCC）组织及白斑中的表达及意义。方法：采用免疫组化Maxvision两步法检测54例口腔鳞状细胞癌、30例口腔黏膜白斑、22例正常口腔黏膜组织中TRAIL的表达水平。结果：TRAIL在口腔鳞状细胞癌组织和黏膜白斑中的表达水平均低于其在正常口腔黏膜中的表达水平（P〈0.05）,分别为33.3%、56.7%、86.4%,其在癌组织与白斑中表达水平的差异无统计学意义。TRAIL在高分化癌组织中的表达水平高于中低分化组（P〈0.05）。TRAIL在不同临床分期组间及淋巴结转移与非转移组间表达水平的差异无统计学意义。结论：TRAIL表达的抑制可能与口腔鳞状细胞癌及口腔黏膜白斑的发生有关,并且TRAIL可能参与了初始阶段的正常黏膜向癌组织的转化。TRAIL在口腔鳞状细胞癌中的表达水平与癌组织的分化程度相关。     

诱导型一氧化氮合酶在口腔黏膜癌前病变——白斑中的表达及意义

目的　探讨口腔黏膜癌前病变 (口腔白斑 )及口腔鳞癌的发生、发展过程中诱导型一氧化氮合酶 (iNOS)的表达及意义。方法　应用免疫组化法对 10例正常口腔黏膜、30例口腔白斑、10例口腔鳞癌分别检测iNOS的表达。结果　iNOS在正常口腔黏膜、口腔白斑、口腔鳞癌的表达阳性率分别为 0、6 6 .7%、80 .0 % ,各组间差异均有非常显著性 (P <0 .0 1)。结论　由iNOS诱导产生的一氧化氮 (NO)可能在口腔鳞癌的发生、发展中起着重要作用。     

Association between proliferative verrucous leukoplakia and infection with human papillomavirus type 16

Proliferative verrucous leukoplakia (PVL) is a recently described clinical entity characterized by multifocal oral lesions that frequently progress to oral cancer despite abstinence from tobacco use by most patients. To determine if this condition is associated with human papillomavirus (HPV), a polymerase chain reaction (PCR) for HPV DNA was performed on 9 lesions from 7 patients with PVL, histologically diagnosed with focal keratosis (1), papilloma (1), epithelial dysplasia (5) and squamous cell cancer (2). Eight (89%) were HPV positive, 7 for HPV 16. For comparison, we studied 55 non-PVL-associated oral specimens, including 24 oral squamous cell cancers. Of the cancers, 8 (33%) were HPV positive, 4 for HPV 16. These data suggest that HPV 16 infection may play an important role in the pathogenesis of PVL-associated oral dysplasia and possibly cancer, but is found in only a small proportion of the more common, non-PVL associated-oral lesions.     

口腔黏膜癌前病变及鳞状细胞癌中VEGF的表达及意义

目的 :探讨VEGF在口腔扁平苔藓、口腔白斑、口腔鳞状细胞癌中的表达及其在口腔鳞癌发生、发展过程中的意义。方法 :应用VEGF多克隆抗体对 10例正常黏膜、2 7例口腔扁平苔藓、2 4例口腔白斑及 30例口腔鳞状细胞癌分别进行免疫组化检测。结果 :VEGF在口腔鳞状细胞癌组织中高表达 ,与口腔扁平苔藓、口腔白斑、正常口腔黏膜组织存在着显著性差异 (P <0 .0 5 ) ;在口腔扁平苔藓和口腔白斑中VEGF有一定的表达 ,且表达强度强于正常口腔黏膜组 (P <0 .0 5 )。结论 :VEGF的过表达与口腔鳞癌的发生、发展有关 ,可以将其作为预测口腔黏膜恶变潜能的重要标志物。     

单层上皮细胞角蛋白在口腔粘膜癌变过程中的表达

目的：探讨单层上皮细胞角蛋白ＣＫ１８和ＣＫ１９作为口腔癌前病变标志的可能性。方法：用ＬＳＡＢ免疫组化染色方法检测ＣＫ１８和ＣＫ１９在福尔马林固定,石蜡包埋的口腔正常粘膜,上皮单纯增生,轻度上皮导演增生,中度上皮异常增生,重度上皮异常增生和口腔鳞癌组织中的分布和表达强度,光镜观察染色切片,结果用秩和检验分析。     

CD147和Ki-67在口腔黏膜白斑和鳞癌组织中的表达及意义

目的：观察CD147和ki-67在口腔正常黏膜、白斑和鳞癌组织中表达的变化,阐明CD147在口腔癌发病机制中的作用。方法：采用免疫组织化学法检测10例正常口腔黏膜、20例白斑伴上皮异常增生上皮和40例鳞癌组织中CD147及Ki-67的表达变化。结果：CD147在正常黏膜阴性表达,白斑和鳞癌组上皮均显著表达CD147;正常组Ki-67表达主要位于上皮棘层,鳞癌组织中Ki-67阳性细胞分布广泛,有大部分侵入固有层中。结论：口腔黏膜发生癌前病变时,即出现CD147表达阳性,随着Ki-67阳性细胞数的增多,癌变细胞增殖不断加强,两者共同作用促进鳞癌的发展。     

ALDH1在OLP和LK组织中的表达及其意义

目的：通过检测干细胞标记物ALDH1在口腔扁平苔藓（OLP）和白斑（LK）中的表达水平,评价其与癌变的关系。方法：采用免疫组织化学SP法检测ALDH1在10例正常口腔黏膜,30例OLP,60例LK,10例口腔鳞癌（OS－CC）中表达水平;再检测ALDH1在30例癌变与30例未癌变LK中的表达差异。结果：ALDH1在正常口腔黏膜中不表达,在OLP、LK和OSCC中的表达率分别为26．7％,63．3％和90．0％（P＜0．05）;ALDH1在未癌变和癌变LK中的表达率分别是43．3％和83．3％（P＜0．01）。结论：ALDH1与口腔黏膜恶性潜能程度相关,可能是预测癌变的分子标记物。     

Human papillomavirus as a risk factor in oral carcinogenesis: a study using in situ hybridization with signal amplification

Introduction:  It is still controversial whether human papillomavirus (HPV) can be considered a risk factor in oral carcinogenesis. The aim of this study was to detect HPV DNA in 50 cases diagnosed as oral leukoplakias, with different degrees of epithelial dysplasia, and as oral squamous cell carcinomas, using in situ hybridization with signal amplification (CSA-ISH).
Methods:  HPV DNA was assessed in paraffin sections using CSA-ISH with a wide-spectrum biotinylated DNA probe. In HPV-positive cases, genotyping with specific probes to HPV types 6/11, 16/18 and 31/33 was performed.
Results:  The overall prevalence of HPV infection was 24%, markedly higher than that found in the control group. Results showed a discrete proportional relationship in the indices found in leukoplakia with no dysplasia, leukoplakia with dysplasia, and squamous cell carcinoma, but this was not statistically significant. When separating the group of leukoplakia by degrees of dysplasia, this relation of proportion was not observed. In genotyping, HPV types 16/18 were the most prevalent, and types 6/11 were only found in groups of mild or no dysplasia.
Conclusion:  The results suggest that HPV is not likely to play a role in the progression of malignant transformation in oral lesions. Nevertheless, the increased prevalence of HPV infection compared to normal oral mucosa and the fact that high-risk HPV types were the most frequently identified do not allow the exclusion of HPV as a risk factor in oral carcinogenesis.     

Nav1.6在口腔黏膜白斑及口腔鳞癌中的表达

目的:探讨钠离子通道Nav1.6在口腔黏膜白斑及口腔鳞癌中的表达及意义.方法:采用免疫组化SP法及Westernblot法检测Nav1.6在口腔正常组织(21例)、白斑(32例)及口腔鳞癌组织(63例)中的表达情况.结果:免疫组化检测,Nav1.6在正常组、口腔白斑组以及癌症组中的阳性表达率分别为6.25％、40％、76.74％;Nav1.6的表达与口腔鳞癌的病理分期有关.Western blot检测,正常组、口腔白斑组、癌症组中Nav1.6的蛋白表达量分别为:0.469±0.015、0.545±0.016、0.848 ±0.020(P ＜0.05).结论:Nav1.6在口腔黏膜癌前病变及口腔鳞状细胞癌的发生和发展中可能起到了一定的作用.     

口腔鳞癌组织中Ki-67和p53蛋白的表达

目的 探讨口腔鳞癌组织中Ki-67和p53蛋白的表达及其与临床病理特征的关系。方法采用免疫组织化学S-P法对10例正常口腔黏膜组织、16例口腔白斑（OLK）组织、48例口腔鳞癌（OSCC）组织中的Ki-67和p53蛋白表达进行检测,结合患者临床病理资料进行分析,使用SPSS17.0 软件包对数据进行统计学处理。结果Ki-67蛋白在正常口腔黏膜组织、口腔白斑和口腔鳞癌组织中的阳性表达率分别为30.0%、56.3%和79.2%;p53的阳性表达率分别为0.0%、43.8%和70.8%,Ki-67和p53在正常黏膜组与口腔白斑和口腔鳞癌组差异均具有显著性（P<0.05）;Ki67蛋白在口腔鳞癌组织中的表达与肿瘤的临床分期、分化程度、有无淋巴结转移有关（P<0.05）,p53蛋白的表达与肿瘤的分化程度有关(P<0.05);Ki-67和p53蛋白在口腔鳞癌组织中的表达呈正相关（P<0.05）。结论Ki-67和p53蛋白在口腔鳞癌组织中高表达,可能在口腔鳞癌的发生、发展过程中起着重要作用。