Studies of the inflammatory process and malignant potential of oral mucosal lichen planus

Oral mucosal lichen planus (OMLP) is a well recognized mucosal disease with unknown aetiology. Considerable controversy exists as to whether OMLP is intrinsically premalignant, or if the disorder facilitates the development of oral mucosal squamous cell carcinoma (OMSCC) by external factors.
The aim of the present studies was to investigate the expression of c-erbB-2 protein in the keratinocytes of the initial biopsies of oral mucosal disorders diagnosed as OMLP with no evidence of epithelial dysplasia and to compare the results with the expression of c-erbB-2 protein in subsequent biopsies obtained from the same patients. These results were compared with the findings from another 26 biopsies from patiente with OMLP and control groups (patiente with dysplasia with no evidence of OMLP, patients with OMSCC with no evidence of OMLP and normal oral mucosa).
The expression of the c-erbB-2 protein was evaluated by Immunohistochemical staining of the gene product with the avidin-biotin-complex method using both fresh frozen and paraffin-embedded tissue sections.
Five of the initial biopsies from patients with OMLP expressed the c-erbB-2 protein and one did not. None of the OMLP cases that subsequently showed evidence of dysplasia expressed the c-erbB-2 protein and of the OMSCC specimens from the patients with OMLP, 2 were negative and 1 expressed c-erbB-2 protein. Within the other group of OMLP specimens 3 (3/26) were negative for c-erbB-2 staining. The specimens from the control groups all expressed the c-erbB-2 protein.
The results indicated the probability of the absence of c-erbB-2 staining being an indication of a potential for neoplastic transformation in OMLP with dysplastic changes.     

Assessment of the value of immunofluorescence microscopy in the diagnosis of oral mucosal lichen planus

Accurate diagnosis of oral mucosal lichen planus (OMLP) is essential if appropriate management is to be instituted. Direct immunofluorescence microscopy (DIF) has facilitated the diagnosis of some dermatologic diseases with oral manifestations, but its use in the diagnosis of OMLP is less well documented. In the present study, 165 biopsies from consecutive cases for which a provisional diagnosis of OMLP was suggested clinically or on the basis of routine histopathologic, or DIF assessment were studied. In 27 cases the results of DIF were non-contributory. In 13 cases the diagnosis could only be established by DIF examination. Nine cases clinically diagnosed as OMLP were assigned to a different condition on the basis of routine histology and DIF. It was concluded that DIF was an integral step in the diagnosis of OMLP.     

The prognosis of oral mucosal squamous cell carcinomas: A comparison of clinical and histopathological grading and of laminin and type IV collagen staining

Changes in the distribution of basement membrane components have been described in dysplastic lesions and in oral mucosal squamous cell carcinomas (OMSCC). The purpose of this study was to determine if these changes were related to pathological grade and if so, whether this had prognostic implications. Fifty formalin-fixed, paraffin-embedded specimens of OMSCC, with five or more years clinical follow-up, were studied using an immunoperoxidase technique for the detection of the basement membrane components, laminin and type IV collagen. The immunoreactivity of each component was evaluated and semiquantitatively scored as minimal, moderate or extensive and the results compared with the tumour size, node involvement and metastasis (TNM) clinical staging system and histopathological features. OMSCC were characterized by minimal or moderate staining with small islands of neoplastic cells frequently lacking staining for laminin and type IV collagen. Deposition of these components decreased with increased histopathological grade and absence of staining was more commonly associated with a poor prognosis. In particular the pattern of type IV collagen staining frequently differed from laminin staining. Neither of these parameters offered an advantage over TNM clinical staging with regard to prognosis. It was concluded that variations in laminin and type IV collagen immunoreactivity occured in OMSCC and that high histopathological grade tumours with considerably diminished staining with anti-laminin and anti-type IV collagen carried a poor prognosis.     

Expression of p53 in oral mucosal hyperplasia, dysplasia and squamous cell carcinoma

OBJECTIVE: To assess p53 expression in a range of oral mucosal lesions and to relate the results to the clinical outcome in patients with dysplastic oral mucosal lesions and oral squamous cell carcinomas (OSCC).
MATERIALS AND METHODS: Archival tissue was available for eight cases of normal oral mucosa, 50 cases of oral mucosal hyperplasia, 41 cases of oral mucosal dysplasia and 48 cases of OSCC. The monoclonal antibody DO-7, reactive to p53 protein, was applied to paraffin-embedded sections using microwave pretreatment and immu-nohistochemical techniques.
RESULTS: The results showed that normal oral mucosa did not express p53.Positive nuclear staining was found in 18/50 (36%) cases of hyperplasia, 35/41 (85%) cases of dysplasia and 45/48 (94%) cases of OSCC.None of the p53 negative dysplasias progressed, while 19% of p53 positive cases of dysplasia recurred following excision and 11% of the cases underwent neoplastic transformation. Five out of 10 (50%) cases of severe dysplasia which were p53 positive resolved.
CONCLUSION: The proportion of cases with positive p53 expression increased from hyperplasia to dysplasia to OSCC. These results may indicate an involvement of p53 in neoplastic transformation as well as in proliferative events although the presence or absence of p53 staining could not be used to predict the outcome of potentially malignant oral mucosal lesions.     

口腔异常增生上皮及鳞癌组织中Fas、FasL及Bcl-2的研究

目的:研究凋亡相关基因Fas、FasL及Bcl-2蛋白在口腔黏膜异常增生上皮及鳞癌组织中变化规律。方法:采用免疫组织化学SP法检测口腔黏膜异常增生上皮及鳞癌组织中凋亡相关基因Fas、FasL及Bcl-2蛋白表达。结果:发现Fas表达水平无差异,但Fas染色在正常、单纯性增生、轻度异常增生组织中以胞膜型为主,而原位癌及鳞癌组织中以胞浆型为主,中度异常增生时两型兼有;FasL在鳞癌组织中过度表达,与对照组、单纯增生组比较差异显著(P<0.05),FasL表达与局部浸润的淋巴细胞呈负相关(r=-0.437,P<0.05)。Bcl-2表达在原位癌时出现高峰,与对照组、单纯增生组比较差异显著(P<0.05),Bcl-2表达水平与Fas L表达水平相关显著(r=0.337,P<0.05)。结论:Fas、FasL及Bcl-2参与了口腔癌前病变癌变发生发展过程,作用的机制可能是它们分别或协同作用抑制细胞凋亡、延长异常细胞生存期、利于免疫逃逸,为细胞癌变提供了条件。     

p53 expression in oral precancer and cancer

Expression of the p53 tumour suppressor gene is a frequent finding in human malignancies, including oral cancer, and it has been detected in some potentially malignant lesions. The results of the present project showed that 35 of the 41 (85 per cent) oral mucosal lesions with histological evidence of epithelial dysplasia expressed p53, but the presence or absence of p53 staining could not be used to predict the outcome of potentially malignant oral mucosal lesions.     

小分子锌指蛋白1在4-硝基喹啉-N-氧化物诱导SD大鼠颊黏膜癌变中的表达研究

目的 研究口腔黏膜癌变中小分子锌指蛋白1（LMO1）在基因转录和蛋白水平的表达变化。方法 取本课题组前期4-硝基喹啉-N-氧化物（4NQO）饮水法构建鳞癌动物模型的49例样本进行苏木精-伊红（HE）染色,依据上皮异常增生程度确定实验组病理分级并确定实验分组;免疫组织化学染色定性确定LMO1的表达部位;实时荧光定量聚合酶链式反应（RT-qPCR）和Western blot检测5组样本中LMO1 mRNA和蛋白的表达。结果 HE染色确定对照组7例,轻度组6例,中度组11例,重度组9例,OSCC组16例。免疫组织化学染色检测可见,LMO1主要表达于细胞质,对照组、轻度组、中度组、重度组和OSCC组的阳性表达率分别为14.3%、33.3%、81.8%、88.9%、93.8%。RT-qPCR检测可见,对照组LMO1 mRNA的表达量最低,OSCC组最高,对照组与轻度组间mRNA表达差异无统计学意义（P>0.05）,其余各组组间两两比较,mRNA的表达差异均有统计学意义（P<0.05）。Western blot检测可见,随着上皮异常增生程度的加重,LMO1的蛋白表达量逐渐上升,在OSCC组中表达最高。结论 口腔黏膜癌变进程中,LMO1 mRNA和蛋白异常表达,且mRNA和蛋白表达量随上皮异常增生程度的加重而增加。     

水通道蛋白3在口腔鳞癌中的表达及临床意义

目的: 探讨水通道蛋白3（AQP3）在口腔鳞癌中的表达及临床意义。方法: 收集2014年3月—2017年4月盘锦辽油宝石花医院保存的202例口腔黏膜组织石蜡标本。其中健康者41例（对照组）、口腔黏膜上皮异常增生45例（A组）、口腔鳞癌116例（B组）。利用免疫组织化学方法检测3组口腔黏膜组织中AQP3的表达情况,分析口腔鳞癌口腔黏膜组织中AQP3表达与临床病理因素及预后的关系。采用SPSS 18.0软件包对数据进行统计学分析。结果: B组口腔黏膜组织中AQP3阳性表达率显著高于A组和对照组（P<0.05）,A组口腔黏膜组织中AQP3阳性表达率显著高于对照组（P<0.05）。肿瘤侵袭深度>5 mm、临床T4分期、颈淋巴结转移、低分化患者,口腔黏膜组织中AQP3阳性表达率分别高于肿瘤侵袭深度≤5 mm、临床T2分期、无颈淋巴结转移、中高分化患者（P<0.05）。AQP3阳性表达的口腔鳞癌患者,术后3年生存率（65.85%）显著低于阴性表达患者（90.00%,P<0.05）。Cox回归分析显示,肿瘤临床分期、分化程度、颈淋巴结转移情况、AQP3阳性表达率是影响总生存率的独立预后因素（P<0.05）。结论: 口腔鳞癌患者口腔黏膜组织中AQP3阳性表达率显著高于口腔黏膜上皮异常增生者,AQP3表达与口腔鳞癌患者临床分期、颈淋巴结转移、分化程度、肿瘤侵袭深度及总生存率相关。     

口腔粘膜癌前病变增殖细胞核抗原表达分布

采用免疫组化染色和图像分析技术，检测了正常口腔粘膜，轻中重度粘膜上皮异常增生和鳞状细胞癌共７４例石蜡包埋组织中增殖细胞核抗原（ＰＣＮＡ）的表达分布．结果显示，正常口腔上皮中，ＰＣＮＡ染色仅见于基层，而所有异常增生和鳞癌标本中，ＰＣＮＡ阳性反应出现在基层以上部位．从轻、中、重度异常增生到鳞癌，ＰＣＮＡ阳性细胞的比率不断增加，每个标记细胞中ＰＣＮＡ染色程度亦显著增强．表明ＰＣＮＡ表达和口腔上皮的增殖潜能及分化程度密切相关，可作为监测口腔粘膜上皮癌变风险的重要生物学标记     

增殖细胞核抗原的表达在口腔粘膜上皮癌变诊断中的意义

目的　研究增殖细胞核抗原 ( PCNA)在口腔粘膜上皮癌变过程中的表达及其在癌变诊断中的意义。方法　采用免疫组化 S-P法对正常口腔粘膜、轻中重度粘膜上皮异常增生和高分化鳞癌共 85例标本进行 PCNA检测。结果　正常口腔粘膜基底细胞层可有少量 PCNA阳性细胞 ;而上皮异常增生和鳞癌时 ,随着病变程度加重 ,PCNA阳性表达也相应提高 ,且阳性反应出现在基层以上部位 ;尤其是重度异常增生 ,其 PCNA阳性表达率较中度异常增生明显增强 ( P=0 .0 0 0 8) ,而重度异常增生与鳞癌间 ,PCNA表达无显著性差异 ( P=0 .72 15 )。结论　 PCNA表达在口腔粘膜上皮癌变诊断中有重要参考价值 ,可作为监测其恶性变的重要生物学指标     

口腔粘膜鳞癌发生过程中Bax蛋白的表达

目的：初步探讨Ｂａｘ基因蛋白产物Ｂａｘ蛋白在口腔粘膜癌前损害发生发展过程中的作用及变化的规律。方法：分别选出正常口腔占膜、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、鳞状细胞癌标本共３８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：光镜下可见,各阶段均有Ｂａｘ蛋白表达,为细胞浆阳性,从正常口腔粘膜组织、上皮单纯增生到中度上皮异常     

Cytokeratin pattern in normal and HPV infected oral mucosa in women with genital HPV infections

The distribution of cytokeratins Nos. 19 (CK 19), 14, 16 and 17 (CK2-27), and 8 and 18 (CK 60-61) in 96 oral mucosal biopsies taken from women with genital HPV infections were studied by immunohistochemistry, using polyclonal antibody CK 19, as well as monoclonal antibodies CK 2-27 and CK 60-61. White staining of the buccal mucosa after acetic acid application, which recently was shown to be affected mostly by smoking and age, could not be explained by differences in cytokeratin pattern. In HPV DNA-positive biopsies, the staining with CK 19 antibody in the basal cell layer was more intense than in HPV DNA-negative biopsies. The staining with CK 2-27 antibody was seen in 76% and 91% of the basal and superficial layers, respectively, even though these low molecular weight cytokeratins should be found mainly from the basal and parabasal cells. CK 60-61 staining was almost similar to that seen recently in normal genital mucosa. When trying to distinguish oral HPV infections from normal mucosa, CK 2-27 and CK 60-61 stainings were of no diagnostic value. The more efficient expression of CK 19 in HPV DNA-positive samples suggests that viral infection might accelerate the production of low molecular weight cytoskeletal protein. This could be interpreted as evidence that HPV might disturb the keratinocyte differentiation in the basal cells. As a result of the present study, CK 19 staining in oral mucosa needs to be further studied in regard to viral infections, because it may help to better understand the interaction between a virus and a host cell.     

The expression of the c-erbB-2 receptor protein in glandular salivary secretions

BACKGROUND: As the maintenance medium of the oral cavity, saliva is secreted from exocrine glands that include the parotid, submandibular, sublingual, and minor salivary glands. Considering that saliva is a fluid suffused with protein, it is possible that the solubilized by-products of oncogenic expression may be present in saliva. Recent studies suggest the presence of solubilized extracellular domain portion of the c-erbB-2 protein in serum, nipple aspirates, and saliva. As a consequence, the purpose of this study was to determine the presence and concentration of c-erbB-2 in major salivary gland secretions. METHODS: Fifteen healthy women had serum, stimulated whole (SWS), parotid (SP), and submandibular/sublingual (SS) salivary secretions collected. The specimens were analyzed for c-erbB-2 using enzyme linked immunosorbent assays (ELISAs). Western blots using c-erbB-2 were also performed on these specimens. RESULTS: The ELISAs revealed the presence of c-erbB-2 in SWS (24.50 Units/ml), SP (19.66 Units/ml), SS (15.59 Units/ml) and serum (1472.15 Units/ml). Western blots confirmed the presence of these 185 kDa proteins. CONCLUSIONS: These results suggest that the protein, c-erbB-2, is present in relatively equal amounts in both SP and SS glandular secretions. Elevated glandular salivary c-erbB-2 concentrations could be useful as a preliminary, non-invasive test in clinical decision making when diagnosing salivary gland carcinomas. Additionally, this marker may have utility in distinguishing between oral lesions that are benign, pre-malignant and malignant in the oral cavity. Further research is required to determine if these findings have clinical utility.     

TGFβ1mRNA在颌骨骨折愈合过程中的表达及意义

为了进一步探讨ＴＧＦβ在骨生长和骨愈合中的作用，建立了颌骨骨折愈合的动物模型，利用原位杂交方法检测骨折愈合过程中ＴＧＦβ１ｍＲＮＡ的表达。结果发现，在骨折愈合过程中的软骨形成和膜内骨骨化阶段，ＴＧＦβ１的转录水平较高，在膜内成骨时转录水平较低，而在伤后即刻反应阶段无ＴＧＦβ１的转录。说明ＴＧＦβ在骨折修复过程中最初的形态发生，随后的细胞增殖及最终的组织重建中起着重要的作用。     

p53 protein expression in sequential biopsies of oral dysplasias and in situ carcinomas

Immunohistochemically detectable levels of p53 may be seen early in the malignant transformation of some neoplasms. To determine if p53 is immunocytochemically detectable, and therefore presumptively abnormal, in oral dysplasias and in situ carcinomas, and to explore the natural history of p53 protein expression in these lesions, sequential biopsies from patients with lesions occurring in the same anatomic site were examined. Formalin-fixed, paraffin-embedded sections from 19 patients were evaluated immunohistochemically for p53 protein using antibody clones Pab1801 and BP53-12. With two exceptions, comparable results were observed with these antibodies. p53 protein was detected immunocytochemically in 6 of 13 patients with dysplasias; 3 of these progressed to p53-positive invasive carcinoma, one advanced to a more severe grade of p53-positive dysplasia, one developed into a p53-negative verrucous carcinoma, and one represented a p53-positive dysplasia developing five years after treatment of a p53-positive carcinoma. The p53-positive dysplasias, which were found in all subtypes (mild, moderate, severe), preceded histologic malignant change by months to years. p53 detection was evident in 4 of 6 patients with in situ lesions. Sequential biopsies of three of these lesions showed no change in lesion histology or p53 staining, and one lesion advanced to a p53-positive carcinoma. It is concluded that p53 protein may be detected early in the development of a subset of p53-positive oral squamous cell carcinomas. This phenomenon may be seen in dysplasias and in situ lesions, and it may have prognostic implications.     

Bcl-2 expression in sequential biopsies of potentially malignant oral mucosal lesions assessed by immunocytochemistry

OBJECTIVE: To examine, for the first time Bcl-2 expression in sequential (autogenous) oral mucosal biopsies taken from the same sites in a gender, risk-factor matched, Caucasoid sample, over a 21-year period. DESIGN: Retrospective immunocytochemical longitudinal study of archival serial biopsies. MATERIALS AND METHODS: Computer records were used to identify biopsy specimens derived from 12 patients. These were divided into four groups: (1) Histologically innocuous lesions which remained histologically innocuous. (2) Dysplastic lesions which remained dysplastic. (3) Histologically innocuous lesions which later progressed to squamous cell carcinoma (SCC). (4) Dysplastic lesions which later progressed to SCC. This represented 65 biopsies in total. Bcl-2 expression was studied using mouse antihuman BCL-2 oncoprotein clone 124 (Dako, Denmark). RESULTS: Generally, there was a lack of Bcl-2 immunoreactivity in the epithelium, with one exception in dysplastic epithelium from a group (3) patient. CONCLUSION: These findings suggest that in our series, Bcl-2 is not expressed early in oral premalignant lesions and appears to contradict previous reports. Possible explanations for this disparity are considered.     

Epithelial growth fraction and expression of p53 tumour suppressor gene in oral submucous fibrosis

The incidence of squamous cell carcinoma in patients with oral submucous fibrosis (OSF) exceeds 7 per cent. The proliferative cell nuclear antigen (PCNA) is a convenient marker of epithelial cell proliferation and p53 tumour suppressor gene mutations or deletions are frequent in oral cancer. The present study estimated the basal epithelial cell growth fraction using a standard immunohistological method for the detection of nuclear PCNA from 20 Nepalese patients with OSF as 31.8 per cent compared with 7.6 per cent for oral mucosa from 43 normal subjects (p<0.001) and 39.4 per cent for 44 patients with oral cancer. The PCNA growth fraction correlated significantly with that derived by Ki-67 labelling. There was no correlation between the growth fraction and the severity of epithelial dysplasia found is OSF.
Abnormal expression of p53 protein identified by immunohistochemistry with a panel of antibodies was found in 70 per cent of the OSF specimens, and 21 per cent of mucosal specimens from subjects with clinically normal mouths. PCNA-positive cells and p53 expression were restricted to the basal epithelial layer in OSF. The unexpected finding of p53 protein in clinically healthy mucosa was confined to subjects aged over 40 years who smoked tobacco, a known risk factor for oral cancer. There was no association between p53 expression and epithelial atypia scores in OSF.
It is concluded that the proportion of actively cycling epithelial cells is increased in OSF and that p53 tumour suppressor gene mutations or deletions may be prevalent. Confirmation by molecular biology techniques of this genetic damage is now needed.     

Shifts in cellular localization of moesin in normal oral epithelium, oral epithelial dysplasia, verrucous carcinoma and oral squamous cell carcinoma

BACKGROUND: Moesin, a member of ERM (ezrin/radixin/moesin) family, links actin filaments of cell surface structure to the cell membrane. The purpose of the study is to assess the shifts in cellular distribution of moesin in normal oral epithelium, oral epithelial dysplasia (OED), verrucous carcinoma (VC), and oral squamous cell carcinoma (OSCC). METHODS: The expression of moesin was evaluated immunohistochemically in paraffin-embedded tissues of 59 specimens of OSCC, 35 specimens of OED, 17 specimens of VC, and five specimens of normal oral epithelium. RESULTS: In the normal oral epithelia, all specimens showed a pattern of membranous expression against the anti-moesin antibody in the basal layer cells. In the OED specimens, moesin was dominantly expressed in the cell membrane except for the cornified layer. In VC and OSCC specimens, almost the whole of the carcinoma cells were stained with anti-moesin antibody. However, in OSCC samples, moesin was markedly expressed increasingly in the cytoplasm and decreasingly in the cell membrane, as compared with OED and VC. In addition, there was a significant correlation between the pattern of moesin expression and tumor differentiation in OSCC. CONCLUSIONS: Our results suggest that it is useful to detect the moesin expression as adjunct to screening mucosal lesions in the oral cavity.     

口腔粘膜上皮癌变过程表皮生长因子受体的表达及意义

目的 研究口腔粘膜上皮癌变过程中表皮生长因子受体的表达及意义。方法 采用免疫组化Ｓ－Ｐ法对正常口腔粘膜,轻中重度粘膜上皮异常增生和高分发化鳞癌共８５例标本进行ＥＧＦＲ检测。结果 正常口腔粘膜全部为阴性。上皮异常增时,随病变程度加重,ＥＧＦＲ阳性表达也相应提高,至重度异常增生时,阳性表达率达最高峰。而高分化鳞癌时,表达率有所下降。结论ＥＧＦＲ的表达与口腔鳞癌的发生有关,可作为评估口腔上皮恶谱潜能的较