P53 and bcl-2 immunoexpression in patients with oral lichen planus and oral squamous cell carcinoma

Objective: The aim of this study was to determine by immunohistochemistry the presence and significance of p53 and bcl-2 proteins in oral lichen planus (OLP) and oral squamous cell carcinoma (OSCC). Study Design: We used 21 cases diagnosed as OLP 16 diagnosed as OSCC and four normal gingival biopsies taken from healthy patients were used as controls. Slides were processed for immunohistochemistry using anti-p53 and anti-bcl-2 monoclonal antibodies. Results: We found p53 immunoexpression in 71.4% OLP cases and 68.7% OSCC cases, with no immunoexpression in control cases. Bcl-2 was negative for all OLP and OSCC cases, and mild positivity was observed in normal tissue. We found significant correlation among p53 expression and OSCC malignancy. Conclusions: Our results suggest that TP53 system mainly promotes a hyperproliferative state by cell cycle arrest of the OLP epithelial cells for repairing damaged DNA nor apoptosis and that anti-apoptotic action of bcl-2 is not important in this disease. Key words:Oral lichen planus, oral squamous cell carcinoma, p53, Bcl-2, carcinogenesis, malignant transformation.     

EP—CAM在OLP和OSCC组织中的表达及意义

目的：观察上皮细胞黏附分子（Ep—CAM）在口腔正常黏膜、扁平苔癣（OLP）和鳞癌（OSCC）组织中表达的变化,分析Ep—CAM在口腔黏膜疾病及其癌变发病机制中的作用。方法：采用免疫组织化学法检测10例正常口腔黏膜、30例OLP（15例非糜烂型、15例糜烂型）及20例OSCC组织中Ep—CAM的表达变化。结果：Ep—CAM在正常黏膜组织阴性表达,而在非糜烂型OLP、糜烂型OLP及OSCC组织中表达逐渐增强。结论：Ep—CAM在口腔黏膜癌变的发展过程中可能具有一定的指示意义。     

Immunohistochemical expression of p53, p16 and hTERT in oral squamous cell carcinoma and potentially malignant disorders

Oral carcinogenesis is a multi-step process. One possible step is the development of potentially malignant disorders known as leukoplakia and erytroplakia. The objective of this study was to use immunohistochemistry to analyze the patterns of expression of the cell-cycle regulatory proteins p53 and p16(INK4a) in potentially malignant disorders (PMD) of the oral mucosa (with varying degrees of dysplasia) and in oral squamous cell carcinomas (OSCC) to correlate them with the expression of telomerase (hTERT). Fifteen PMD and 30 OSCC tissue samples were analyzed. Additionally, 5 cases of oral epithelial hyperplasia (OEH) were added to analyze clinically altered mucosa presenting as histological hyperplasia without dysplasia. p53 positivity was observed in 93.3% of PMD, in 63.3% of OSCC and in 80% of OEH. Although there was no correlation between p53 expression and the grade of dysplasia, all cases with severe dysplasia presented p53 suprabasal immunoexpression. p16(INK4a) expression was observed in 26.7% of PMD, in 43.3% of OSCC and in 2 cases of OEH. The p16(INK4a) expression in OEH, PMD and OSCC was unable to differentiate non-dysplastic from dysplastic oral epithelium. hTERT positivity was observed in all samples of OEH and PMD and in 90% of OSCC. The high hTERT immunoexpression in all three lesions indicates that telomerase is present in clinically altered oral mucosa but does not differentiate hyperplastic from dysplastic oral epithelium. In PMD of the oral mucosa, the p53 immunoexpression changes according to the degree of dysplasia by mechanisms independent of p16(INK4a) and hTERT.     

血管内皮生长因子在口腔扁平苔藓及鳞癌组织中的表达

目的探讨血管内皮生长因子在口腔扁平苔藓癌变及发病机制中的作用。方法采用免疫组化法,检测10例正常口腔黏膜、25例口腔扁平苔藓、11例口腔扁平苔藓伴不典型增生及14例口腔鳞癌,上皮组织中血管内皮生长因子的表达水平。结果血管内皮生长因子在24例口腔扁平苔藓标本中阳性表达2例,阴性表达22例,阳性表达率明显低于其它3组（P〈0.05）。扁平苔藓伴不典型增生及口腔鳞癌中血管内皮生长因子的阳性表达率均高于正常黏膜（P〈0.05）。结论血管内皮生长因子的表达异常可能在口腔扁平苔藓的发生发展及癌变的过程中起作用。     

Oral lichen planus has a high rate of TP53 mutations. A study of oral mucosa in icelanD

Oral squamous cell carcinoma (OSCC) is a world-wide health problem. In addition to external exposure (smoking and alcohol), certain oral lesions may increase the risk of oral cancer (e.g. leukoplakia, erythroplakia, and oral lichen planus). TP53 has been implicated in OSCC, but there are limited studies of mutations in premalignant oral lesions. In this study, 55 samples from OSCC, 47 from hyperkeratotic (HK) oral mucosa, clinically diagnosed as white patches, 48 samples from oral lichen planus (OLP), and 12 biopsies from normal oral mucosa were studied immunohistochemically for expression of TP53 protein. From all the carcinoma samples and selected non-malignant samples showing moderate or strong TP53 protein expression, malignant cells or TP53-positive nuclei were microdissected and screened for mutations in exons 5-8 by constant denaturation gel electrophoresis. Moderate to strong TP53 protein staining was seen in 56% of OSCC, 32% of OLP but only in 13% of HK. All OLP samples showed a characteristic pattern of positive nuclei confined to the basal layer, whereas TP53 staining was seen in suprabasal nuclei in HK. Mutation rate was 11 out of 52 for OSCC, three out of 20 tested for HK and, remarkably, nine out 27 tested for OLP. There was no correlation between TP53 protein staining and TP53 mutations. No associations were found with anatomical sites or disease progression. The unexpectedly high mutation rate of OLP might explain the premalignant potential of this lesion.     

口腔鳞癌中HPV感染及其对p5 3改变影响的研究

目的：探讨高危型人乳头状瘤病毒（ＨＰＶ）在口腔鳞癌中的感染情况及其对Ｐ５３蛋白表达和ｐ５３突变的影响。方法：采用免疫组化和ＰＣＲ－ＳＳＣＰ方法，分别检测４０例来癌中高危型ＨＰＶＥ６蛋白表达、Ｐ５３蛋白表达和ｐ５３基因突变的情况。结果：９例ＨＰＶＥ６蛋白染色阳性，阳性率２２．５％（９／４０），与正常粘膜对照组有显著差异（Ｐ＝０．０２１）。ＨＰＶ阳性组中Ｐ５３蛋白表达率１１．１％（１／９），ＨＰＶ阴性     

Prognostic impact of p53 and p63 immunoexpression in oral squamous cell carcinoma

BACKGROUND: The role of p53 and p63 proteins in the prognosis of oral squamous cell carcinoma (OSCC) is still debatable. Our aim here was to investigate the relationship between the immunoexpression of these proteins with some clinicopathologic parameters of prognostic significance in OSCC. METHODS: Formalin-fixed paraffin-embedded sections from 106 patients were used for study together with the following data: primary site, histologic differentiation, recurrences, metastasis, disease-free survival and overall survival (OS). RESULTS: In OSCCs, the positive rate for p63 protein immunoexpression (87.8%) was higher than p53 (52.8%). p53 expression correlated with metastasis. Tumors negative for p53 and with strong intensity for p63 expression had a significantly higher OS. CONCLUSIONS: p53 overexpression is associated with a larger number of metastases and is correlated with a poor outcome as well as decreased intensity in p63 immunoexpression.     

iNOS在口腔扁平苔藓癌变过程中的表达研究

目的 :研究诱导型一氧化氮合酶 (iNOS)在口腔扁平苔藓、口腔鳞状细胞癌发展过程中的表达水平及其作用。方法 :以正常口腔黏膜作对照 ,采用免疫组化SP法检测 2 5例口腔扁平苔藓和 10例口腔鳞状细胞癌组织中的iNOS表达。结果 :正常口腔黏膜iNOS阴性表达 ;扁平苔藓组和鳞癌组iNOS表达较正常黏膜组均非常显著增加 (P <0 .0 1) ;扁平苔藓组与鳞癌组之间以及扁平苔藓组内部按上皮异常增生程度分组之间iNOS的表达均无显著差异 (P >0 .0 5 )。结论 :iNOS在口腔扁平苔藓伴上皮异常增生和鳞癌中高表达 ,在口腔鳞癌的发生发展中起着重要的作用。     

口腔扁平苔藓患者细胞凋亡及凋亡相关基因p53,bcl—2,C—myc的表达

目的研究口腔扁平苔藓(OLP)患者病变组织中细胞凋亡的发生情况及凋亡相关基因p53、bcl-2、C-myc的表达,探讨其在 OLP病变发生中的意义.方法采用TUNEL法及免疫组织化学法,检测45例OLP(糜烂型23例,非糜烂型22例)中凋亡细胞情况,计算凋亡指数(AI)及p53、bcl-2、C-myc的表达情况. 结果 45例OLP均有凋亡细胞发生,AI为24.3%±12.0%(1.9%～50.5%);其中糜烂型AI为19.0%±10.4%(1.9%～ 50.5%),非糜烂型AI为29.4%±11.2%(10.4%～49.3%),两者间有显著差异(P＜0.05).OLP组织中p53、bcl-2、C-myc表达阳性率分别为31.1%、35.6和33.3%.其中糜烂型OLP中的p53、bcl -2、C-myc表达阳性率分别为39.1%、52.2%和43.5%,非糜烂型阳性率分别为22.7%、18 .2%和22.7%,两组间仅bcl-2的表达有显著差异(P＜0.05).结论细胞凋亡及凋亡相关基因p53、bcl-2、C-myc参与了OLP病变的发生发展,AI的变化与病变部位的慢性炎症有关,bcl-2在糜烂型OLP中的表达应引起高度重视.     

p53-p21waf1通路在口腔鳞状细胞癌中心体扩增中的作用及意义

目的 了解口腔鳞状细胞癌(oral squamous cell cancer,OSCC)p21waf1蛋白表达与中心体扩增的相关性,探讨p53-p21waf1通路在OSCC中心体扩增中的作用及意义.方法 8例正常口腔黏膜及27例OSCC石蜡包埋组织,采用间接免疫荧光双重染色法了解OSCC组织中心体扩增情况;采用流式细胞术及免疫组织化学方法对相应组织p21waf1蛋白表达量及突变型p53蛋白进行检测,分析三者间的相关性.结果 中心体数目扩增(＞2个/细胞)可见于78%(21/27)的OSCC组织,p21waf1蛋白表达量在有中心体扩增的OSCC组织中[(0.878±0.081)]低于无中心体扩增的OSCC组织[(0.952±0.018),t=3.838,P=0.001],OSCC组织中心体扩增程度与p21waf1蛋白表达量间存在负相关关系(r=0.472,P＜0.05);p21waf1蛋白表达量在OSCC组织p53阳性组[(0.823±0.071)]低于p53阴性组[(0.909±0.075),t=3.905,P＜0.01],两者间存在负相关关系(r=-0.491,P＜0.05).结论 p53-p21waf1通路可能参与了OSCC中心体循环调控,p53突变导致的p21waf1蛋白表达下调在OSCC中心体扩增中可能起一定作用.     

p63在口腔扁平苔藓中的表达

目的探讨p53家族新成员p63在口腔扁平苔藓（OLP）中的表达，其与OLP发生、发展的关系以及是否可作为一种标志物预测OLP的转归。方法采用免疫组织化学方法检测30例012中p63蛋白的表达，同时采用RT—PCR方法分析p63编码的两大类转录产物TAp63和ANp63的mRNA水平，并且与口腔鳞状细胞癌（OSCC）及正常口腔黏膜（NOM）进行比较。结果p63蛋白在NOM、OLP和OSCC组中均有表达，与NOM比较，其累积光密度（IOD）值在OLP中下调，在OSCC中明显上调。在糜烂萎缩型OLP中，p63蛋白表达高于非糜烂型（P〈0．001）。RT-PCR检测结果显示，TAp63的mRNA水平在OLP中表现为中等。略低于NOM．明显高于OSCC；ANp63的mRNA水平在NOM和OLP中极低．其转录扩增带大多数检测不到．但在OSCC组都能检测到．而且呈高水平表达。TAp63的mRNA水平在糜烂萎缩型OLP中低于非糜烂型（P=0．018），ANp63则高于非糜烂型（P=0．049）。结论p63与OLP的发病机制有关，其中TAp63和ANp63分别起不同的作用。p63表达的高低可能可以作为预测OLP是否有高风险癌变的检测指标。     

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.     

端粒酶逆转录酶mRNA在OLP和OSCC中的表达

目的：探讨端粒酶逆转录酶（TERT）mRNA在口腔扁平苔藓（OLP）和口腔鳞状细胞癌（OSCC）中的表达以及在OLP癌变过程中的作用。方法：应用mRNA原位杂交法对正常口腔黏膜12例,非糜烂型OLP20例,糜烂型OLP20例及OSCC20例,进行hTERTmRNA的检测。结果：正常口腔黏膜、非糜烂型OLP、糜烂型OLP及OSCC中hTERTmRNA阳性表达率分别为8.33％（1／12）、15％（3／20）、45％（9／20）、80％（16／20）。其中,除正常口腔黏膜与非糜烂型OLP中hTERT mRNA阳性率无显著性差异外,其余各组间均有显著性差异（P〈0.05）。而且,正常口腔黏膜与非糜烂型OLP的hTERT mRNA染色强度明显低于糜烂型OLP及OSCC（P〈0.05）。结论：hTERT mRNA可能在糜烂型OLP的癌变过程起着一定的作用。     

Role of TP53 in the progression of pre-malignant and malignant oral mucosal lesions. A follow-up study of 144 patients

Background:  Prediction of progression from pre-malignant oral mucosal lesions to malignancy, or recurrence of an existing oral squamous cell carcinoma (OSCC), is an important clinical problem in oral medicine.
Methods:  This study presents a follow-up of a study published in 2002. Samples from 54 patients with OSCC, 45 with oral lichen planus (OLP) and 45 with hyperkeratosis (clinically leukoplakia), diagnosed between 1987 and 1996, were analysed for TP53 protein expression and TP53 mutation. Follow-up was 11–17 years for OSCC (mean 13.3), 12–22 years for OLP (mean 15.9) and 12–17 years for hyperkeratosis (mean 14.5).
Results:  Of the 54 OSCC patients, 28 experienced recurrent disease, 21 died of OSCC, 22 died of other causes. Of the 14 OSCC patients with mutated TP53 ( n  = 11), the cancer recurred in eight (57%) and in 20/39 (51%) without mutation. Expression of TP53 protein was significantly associated with reduced overall survival. Among OLP patients, nine were TP53- mutated out of 31 tested. One TP53- mutated OLP patient developed OSCC in a different site. Of the hyperkeratosis patients, three were mutated of 22 tested. One hyperkeratosis patient (non-mutated) developed OSCC in the same site.
Conclusion:  TP53 mutations can exist in benign oral mucosal lesions for many years without progression to malignancy. No association was found between TP53 protein expression or TP53 mutation and recurrence of OSCC or disease-related survival. Overall survival was reduced in patients with positive TP53 protein expression.     

口腔黏膜癌前病变和口腔鳞状细胞癌中Stat3的表达及意义

目的研究口腔黏膜癌前病变和口腔鳞状细胞癌(OSCC)中细胞信号传导和转录激活因子(Stat3)的表达及意义。方法采用免疫组化方法分别检测9例正常口腔黏膜,口腔扁平苔藓(OLP)和白斑(OLK)共8例,OLP、OLK伴异常增生22例,OSCC19例中Stat3的表达。结果Stat3阳性表达分布于细胞质和细胞核内。正常口腔黏膜、单纯增生、异常增生和OSCC中Stat3阳性表达率分别为11.11%(1/9)、12.50%(1/8)、59.09%(13/22)和84.21%(16/19)。OSCC与正常口腔黏膜、单纯增生、异常增生相比,差异有显著性(P<0.05);异常增生与正常口腔黏膜、单纯增生相比,差异有显著性(P<0.05);而单纯增生和正常口腔黏膜相比差异无显著性(P>0.05)。结论Stat3与口腔黏膜癌变的发生发展有着密切关系,对Stat3表达的研究将有助于口腔黏膜癌前病变癌变的检测和OSCC的早期诊断。     

P53和Mdm2蛋白在人口腔扁平苔藓和口腔鳞状细胞癌中的表达

目的：探讨正常口腔黏膜组织（NOM）、口腔扁平苔藓（OLP）、口腔鳞状细胞癌（OSCC）组织中,抑癌基因P53和癌基因Mdm2的表达情况及关系。方法：采用免疫组化SP法检测12例正常口腔黏膜组织、40例OLP组织、33例OSCC组织中P53、Mdm2基因的表达情况,并分析其相关性。结果：P53蛋白在NOM、OLP及OSCC组织中的阳性表达率呈明显的逐渐升高趋势。Mdm2蛋白在NOM中不表达,在OLP中呈现弱阳性表达,在OSCC不同病理分级中阳性表达率依次增高。各组表达均有统计学意义（P〈0.05）。另外P53蛋白、Mdm2蛋白在OS-CC组织中的表达关系无明显的相关性。结论：P53、Mdm2蛋白的表达与OLP及OSCC的发生、发展密切相关。     

口腔鳞状细胞癌组织中P53、bcl-2、nm23蛋白表达与肿瘤生物学行为的关系

目的观察P53,bcl-2,nm23蛋白在口腔鳞状细胞癌(鳞癌)组织中的表达情况,以探讨其在口腔鳞癌生物学行为及预后预测方面的意义。方法采用免疫组化S-P法检测65例口腔鳞形细胞癌(OSCC)组织中P53,bcl-2,nm23蛋白的表达,分析其与肿瘤病理表现的相关性。结果在65例口腔鳞癌中,P53,nm23,bcl-2蛋白阳性检出率分别为56.92%,53.85%,46.15%。P53蛋白阳性表达率与各项病理参数均无明确相关关系,但表达水平随肿瘤分化程度降低而增高;bcl-2蛋白阳性表达率与肿瘤组织学分级和浸润方式有关(P<0.05),中低分化和微灶浸润性生长的肿瘤有较高的阳性表达率;nm23蛋白阳性表达率与颈淋巴结转移明显负相关(P<0.01)。结论P53,nm23,bcl-2蛋白表达与口腔鳞癌的生物学行为关系密切,有助于评估肿瘤进展和预后预测。     

Prevalence of Epstein-Barr virus in oral squamous cell carcinoma,oral lichen planus,and normal oral mucosa

OBJECTIVE: The Epstein-Barr virus (EBV) is associated with both malignant and benign diseases in the head and neck region. In several studies it has also been associated with oral squamous cell carcinoma (OSCC). Oral lichen planus (OLP) is a disease with unknown origin, and viral antigens have been proposed as etiologic agents. Smoking and alcohol habits are known risk factors for oral cancer development. In this study, the prevalence of EBV in OSCC and OLP was investigated, along with the effect of smoking, alcohol use, and age on EBV prevalence. STUDY DESIGN: We examined 29 patients with OSCC, 23 with OLP, and 67 with clinically healthy oral mucosa. For EBV DNA analysis, a nested polymerase chain reaction method was used. RESULTS: The overall EBV prevalence in patients with oral disease was 32.1%. Of the OSCC patients, 37.9% were EBV positive; and of the OLP patients, 26.1% were EBV positive. Both percentages were statistically significant compared with that of control patients (7.3%). The difference in EBV prevalence between the smoking control group and the nonsmoking control group was insignificant. Increased age did not enhance EBV prevalence. CONCLUSIONS: This investigation shows that EBV is present in oral diseases such as OSCC and OLP. Smoking, alcohol use, or age does not seem to be a risk factor for EBV infection. The etiologic role of EBV in OSCC and OLP needs to be examined in a prospective follow-up study.     

口腔黏膜癌变过程中丝氨酸/苏氨酸激酶15的表达及意义

目的了解丝氨酸/苏氨酸激酶15（STK15）在口腔黏膜癌变过程中的表达变化,探讨P53/STK15转激活-非依赖通路在口腔鳞癌（OSCC）发生发展中的作用及意义。方法正常口腔黏膜8例,上皮异常增生患者27例,OSCC患者43例, 石蜡包埋组织,采用免疫组化SABC法了解STK15及P53蛋白表达情况,分析二者的相关性及其临床病理学意义。结果STK15在正常口腔黏膜无表达,在上皮异常增生及OSCC中阳性率分别为40.74%（11/27） 和67.44%（29/43）,各组间差异均有统计学意义（P<0.05）;口腔鳞癌中STK15阳性率在P53阳性组高于P53阴性组,在OSCC有淋巴结转移组高于无淋巴结转移组,差异均有统计学意义（P<0.05）。结论STK15过表达是口腔黏膜癌变过程的早期事件,口腔鳞癌STK15过表达可能与p53突变有关并与OSCC淋巴结转移密切相关,P53/STK15转激活-非依赖通路在OSCC发生发展中可能起重要作用。