p16,p53,Ki67在口腔癌前病变表达及4年临床随访

目的 :研究 p16,p5 3 ,Ki-67蛋白表达与口腔癌前病变的关系。 方法 :采用免疫组织化学LsAB法对43例上皮异常增生 ( 2 0例轻度上皮异常增生 ,2 3例重度上皮异常增生 )和 2 0例正常口腔黏膜 p16,P5 3和Ki -67蛋白的表达进行研究 ,并对上皮异常增生患者实际癌变率做了 4年追踪。结果 :正常黏膜组 ,p5 3不表达 ,Ki -67少量表达 ,p16的阳性表达为 10 0 %。轻度上皮异常增生 ,p5 3 ,Ki-67少数表达 ,p16表达率为 86.96%。重度上皮异常增生 ,p5 3和Ki-67过度表达 ,p16表达明显下降 ,与正常黏膜 ,轻度上皮异常增生相比差异显著 (P <0 .0 5 )。p5 3和Ki-67蛋白过度表达而 p16呈低表达与实际口腔癌前病变癌变率有一定相关性。 结论 :口腔黏膜癌变是一个由量变到质变的过程 ,它们的调控基因 p16,p5 3 ,Ki -6发生了显著的变化 ,可能起着重要的调控作用。     

口腔白斑癌变过程中整合素β1的表达及其与细胞增殖的关系

目的 探讨整合素β1在口腔白斑癌变发生、发展中的可能作用及其与细胞增殖的关系.方法 免疫组化SP法研究整合素β1及Ki-67在12例正常口腔黏膜、10例单纯增生白斑、24例异常增生白斑、14例早期浸润癌组织中的表达情况及关系.结果 在正常黏膜及单纯增生白斑中整合素β1表达于基底及副基底层细胞的胞膜;Ki-67在基底及副基底层的胞核中表达.在异常增生白斑及早期浸润癌中可见整合素β1表达于基底层、增生的棘层细胞及癌细胞胞膜;Ki-67表达于基底层、增生的棘层细胞及癌细胞的胞核.整合素β1在异常增生及早期浸润癌中的强阳性表达率分别为29%(7/24)、57%(8/14);Ki-67在异常增生及早期浸润癌中的强阳性表达率分别为42%(10/24)、57%(8/14).整合素β1、Ki-67在4组病变之间的阳性分级差异有统计学意义(χ2=10.651,P=0.014;χ2=14.831,P=0.002);整合素β1在正常黏膜、单纯增生组与早期浸润癌组之间的差异有统计学意义(P=0.008,P=0.013).Ki-67在正常黏膜组与异常增生组、早期浸润癌组之间差异有统计学意义(P=0.026,P=0.001),单纯增生组与早期浸润癌组之间差异有统计学意义(P=0.005).整合素β1与Ki-67的表达显著相关(R=0.442,P＜0.01).结论 整合素β1在口腔白斑异常增生及早期浸润癌中表达增强,可能与促进细胞增殖有关.     

Immunohistochemical expression of EGFR in oral leukoplakia: Association with clinicopathological features and cellular proliferation

Objectives: to investigate the immunoexpression of epidermal growth factor receptor (EGFR) in a sample of oral leukoplakias (OL) and to determine the receptor’s association with dysplasia, tobacco consumption, lesion site, and proliferation rate. Although EGFR should be overexpressed in some oral leukoplakias, the factors that may interfere with this expression and the influence of this receptor on epithelial proliferation have yet to be investigated. Study Design: Samples of oral leukoplakias (48) and of normal oral epithelium (10) were immunohistologically examined for expression of EGFR. Immunohistochemistry for Ki-67, and p27 were also performed in leukoplakias. EGFR expression was associated with clinical and pathological features. Results: EGFR was positive in 62.5% of the leukoplakias and 50% of normal oral epithelium. The number of EGFR positive OL located in high-risk sites was significantly higher than EGFR positive OL located in low-risk sites. Most of the p27 negative leukoplakias were EGFR positive, and the p27 index in the parabasal layer was diminished in the presence of dysplasia. Positivity for EGFR was not associated with dysplasia, tobacco exposure, or Ki-67. Conclusion: EGFR is expressed in leukoplakia regardless of dysplasia, but EGFR positivity should be more frequent in lesions sited in areas of high cancer risk. The association between EGFR and p27 may represent an important mechanism in the control of cellular proliferation and malignant progression of oral epithelium and therefore warrants further investigation. Key words:Oral leukoplakia, EGFR, p27, Ki-67, epithelial dysplasia.     

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。     

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.     

Maspin, p53, p63, and Ki-67 in epithelial lesions of the tongue: from hyperplasia through dysplasia to carcinoma

Background:  The pattern of changes in the expression of ma mmary s erine p rotease in hibitor (maspin) tumor suppressor protein in tongue epithelial lesions [hyperplasia (HP), mild dysplasia (MD), moderate-to-severe dysplasia (MSD) and squamous cell carcinoma (SCC)] was investigated and correlated to the expression of maspin-regulating factors p53 and p63, and the proliferation marker Ki-67.
Methods:  Cases of HP ( n  = 16), MD ( n  = 12), MSD ( n  = 11), and SCC ( n  = 22) were immunostained for maspin, p53, p63, and Ki-67. Maspin expression was scored separately for the basal, middle, and upper thirds of the epithelial width, and as the total sum of all 'thirds' (maspin-total). p53, p63, and Ki-67 were immuno-morphometrically assessed for the entire epithelial width.
Results:  Maspin expression was differential and progressive extending to higher epithelial layers as dysplastic changes aggravated and culminated in carcinoma. Strong expression was related to MSD in the middle third and to carcinoma in the upper third. It was frequently lost at the invasion front, where the tumor was less differentiated. The changes in mean scores of maspin-total in the different study groups were positively correlated to the mean scores of p63 ( r  = 0.5, P  <   0.001), p53 ( r  = 0.4, P  =   0.004), and Ki-67 ( r  = 0.5, P  <   0.001).
Conclusions:  Strong expression of maspin in the middle third of the epithelium may be considered a diagnostic sign of mild-to-moderate dysplasia and an indication of carcinoma in the upper third. The correlations between maspin and controlling factors (e.g. p63 and p53) may be events with key roles in the development of tongue carcinoma.     

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。     

Immunohistochemical expression of EGFR in oral leukoplakia: Association with clinicopathological features and cellular proliferation

Objectives: to investigate the immunoexpression of epidermal growth factor receptor (EGFR) in a sample of oral leukoplakias (OL) and to determine the receptor' s association with dysplasia, tobacco consumption, lesion site, and proliferation rate. Although EGFR should be overexpressed in some oral leukoplakias, the factors that may interfere with this expression and the influence of this receptor on epithelial proliferation have yet to be investigated. Study Design: Samples of oral leukoplakias (48) and of normal oral epithelium (10) were immunohistologically examined for expression of EGFR. Immunohistochemistry for Ki-67, and p27 were also performed in leukoplakias. EGFR expression was associated with clinical and pathological features. Results: EGFR was positive in 62.5% of the leukoplakias and 50% of normal oral epithelium. The number of EGFR positive OL located in high-risk sites was significantly higher than EGFR positive OL located in low-risk sites. Most of the p27 negative leukoplakias were EGFR positive, and the p27 index in the parabasal layer was diminished in the presence of dysplasia. Positivity for EGFR was not associated with dysplasia, tobacco exposure, or Ki-67. Conclusion: EGFR is expressed in leukoplakia regardless of dysplasia, but EGFR positivity should be more frequent in lesions sited in areas of high cancer risk. The association between EGFR and p27 may represent an important mechanism in the control of cellular proliferation and malignant progression of oral epithelium and therefore warrants further investigation.     

Reduction of syndecan-1 expression is associated with dysplastic oral epithelium

Syndecans are a family of integral membrane proteoglycans that participate in cell-matrix interactions and growth factor binding. Syndecan-1 expression is induced during keratinocyte differentiation and reduced in squamous cell carcinomas. The purpose of this study was to examine the alteration in syndecan-1 expression in dysplastic oral epithelium. Sixty-six oral biopsy specimens (43 epithelial dysplasias, 3 carcinoma in situ and 20 squamous cell carcinomas) were studied using immunohistochemical methods. The normal epithelium specimens were highly positive for syndecan-1. Fifteen of 46 dysplasias or carcinoma in situ specimens showed negative or weak staining for syndecan-1, two of which were totally negative. Intermediate and strong staining were observed in 17 and 14 dysplasias or carcinoma in situ specimens, respectively. Thirteen (65%) squamous cell carcinomas showed negative or weak staining for syndecan-1, seven of which were totally negative. Only three carcinomas had a strong syndecan-1 expression. Four of the 34 patients with dysplasia who were followed up developed squamous cell carcinoma. All these dysplasias had weak or totally negative syndecan-1 expression. The results suggest that the loss of syndecan-1 is associated with dysplastic changes in oral epithelium.     

Prevalence of p53, bcl-2, and Ki-67 immunoreactivity and of apoptosis in normal oral epithelium and in premalignant and malignant lesions of the oral cavity.

PURPOSE: Loss of normal p53 is correlated to the progression of several preneoplastic lesions to neoplasms, and overexpression of bcl-2 determines an alteration of programmed cell death. There is an increased awareness of the importance of apoptosis in cancerogenesis, and a strong correlation of Ki-67 with high tumor grade has been reported. MATERIALS AND METHODS: The aim of our study was to investigate immunohistochemically the expression and relationship of p53, bcl-2, MIB-1, and the apoptotic index (AI) in normal oral epithelium, leukoplakia, dysplasia, and oral squamous cell carcinoma. RESULTS: A strong correlation was found between p53 overexpression and cell proliferation (MIB-1) and the AI. An inverse relationship was found between bcl-2 expression and MIB-1 and AI. A significant inverse relationship was found between p53 and bcl-2. A good positive correlation was present between AI and MIB-1 expression. CONCLUSIONS: Apoptosis could be important to help to understand oral carcinogenesis.     

Histomorphometry and immunohistochemical features of grade I (WHO) oral radiomucositis

AIMS: The aims of this study were to describe the immunohistopathological and morphometric features of oral mucositis grade I (WHO). MATERIAL AND METHODS: Ten samples of oral mucositis were biopsied and submitted to histopathological, morphometric and immunohistochemical analyses (CD68, Ki-67 and p53). The samples were compared with the buccal mucosa of head and neck cancer patients before radiotherapy (NMCP), normal buccal mucosa (NM) and oral dysplasia (OD). RESULTS: Epithelial thickness, area and perimeter were decreased in oral mucositis and inflammatory components, increased when compared with NMCP. CD68 immunoreactivity, near to the epithelium, was more evident in oral mucositis than in NMCP (P = 0.01). The Ki-67 counts were higher in oral mucositis than in NM and NMCP (P = 0.001 and P = 0.043, respectively), but without any difference with OD (P = 0.284). The p53 staining was present in all cases of mucositis and oral dysplasia, but negative in NMCP and NM. CONCLUSIONS: Oral mucositis grade I (WHO) presented epithelial atypia and atrophy, increased inflammatory response, with relevant Ki-67 count and positiveness for p53.     

艾滋病感染者口腔疣状肿块人类乳头状瘤病毒感染与P53,Ki-67蛋白过度表达

目的 :了解艾滋病感染者口腔黏膜疣状肿块细胞生物学特性。方法 :应用免疫组织化学法、PCR对艾滋病感染者口腔黏膜疣状肿块、非艾滋病感染者口腔黏膜癌前病变和鳞癌组织中P5 3和Ki 67蛋白、人类乳头状瘤病毒 (HPV ) ,巨细胞病毒 (CMV)和EB病毒 (EBV )进行检测。结果 :( 1)艾滋病感染者口腔黏膜疣状肿块中P5 3蛋白阳性表达率为 2 3 % ,Ki 67蛋白阳性表达率为 76% ,二者均低于非艾滋病感染者口腔黏膜鳞癌(P <0 .0 5 ) ,但与非艾滋病感染者口腔黏膜癌前病变无明显差别 (P >0 .0 5 ) ;( 2 )艾滋病感染者口腔黏膜疣状肿块中 ,HPV感染率为 88.2 % ,明显高与非艾滋病感染者口腔黏膜癌前病变和口腔黏膜鳞癌 (P <0 .0 1)。没有检测到EBV、CMV病毒感染。结论 :艾滋病感染者口腔疣状肿块和HPV感染有关 ,存在抑癌基因突变和细胞过度增殖现象。     

The presence of p53 protein in relation to Ki-67 as cellular proliferation marker in head and neck squamous cell carcinoma and adjacent dysplastic mucosa

Paraffin embedded material from 15 patients suffering from head and neck squamous cell carcinoma (NNSCC) bordered by dysplastic mucosal areas was immunohistochemically investigated for the presence of p53 protein and Ki-67 proliferation marker. p53 protein was present in 9 cases (60%), invariably in invasive cancer areas as well as in adjacent non-invasive dysplastic mucosa. Only cells exhibiting atypia contained p53 protein. Ki-67 proliferation marker was present in the basal cells of the normal epithelium and more extensive in dysplasias and HNSCC. The presence of Ki-67 closely coincided with p53 protein in the 9 cases exhibiting this. No differences in Ki-67 expression were found between p53 positive and negative cases. It is concluded that the appearance of p53 protein occurs early in carcinogenesis but that cells also may show increased proliferation without involving immunohistochemically detectable alterations in the p53 gene.     

Ⅳ型胶原和C—erbB—2在口腔念珠菌白斑中的表达

目的 研究口腔念珠菌白斑Ⅳ型朱和癌基因蛋白Ｃ－ｅｒｂＢ－２的表达特点。方法 应用ＳＰ免疫组织化学染色法,对３２例口腔念珠菌白斑病例中的Ⅳ型胶原和癌基因蛋白Ｃ－ｅｒｂＢ－２的表达特点进行分析。结果 Ⅳ型胶原染色在２７例中成功。正常和单纯境生的口腔上皮苦底膜区Ⅳ型胶原呈均匀线状阳性,发生改变时表现为增厚、变薄和染色中断。上皮异常增生时变薄和染色中断者多于非上皮异常增生者。Ｃ－ｅｒｂＢ－２在７８％的念珠     

Assessment of c-Jun, c-Fos and cyclin D1 in premalignant and malignant oral lesions

Some oral cancers are known to develop from dysplastic oral epithelium. In the present study, the expression of c-Jun, c-Fos, and cyclin D1 proteins in oral epithelial lesions with different degrees of dysplasia, and in oral squamous cell carcinomas (OSCCs) was evaluated. Eighteen cases of mild dysplasia, 23 cases of moderate to severe dysplasia and 24 OSCCs were studied immunohistochemically. Additionally, 15 sections of oral mucosa without any evidence of dysplasia were included in the study. Results: c-Jun expression increased according to the degree of oral dysplasia, with the greatest expression found in OSCC. c-Fos expression was intense in normal mucosa, reduced in mild dysplasia and high in moderate to severe dysplasia and in OSCCs. Cyclin D1 was expressed in only a few cases of moderate to severe dysplasia and in most of the OSCCs. Statistical analysis showed a correlation between the three proteins and the degree of epithelial alteration. The present results indicate a possible role of c-Jun and c-Fos in malignant transformation of oral mucosa.     

Immunohistochemical analysis of the p53 tumor suppressor gene product in oral leukoplakia

Squamous Cell Carcinoma (SCC) is the most frequent malignancy in the oral cavity. p53 protein has been reported to be expressed at high levels in malignant lesions, while the level in premalignant lesions has yet to be determined. In this study, oral leukoplakia and oral SCC were examined. Seventy-four incision or excision samples from 43 cases diagnosed as leukoplakia, and 41 samples from 37 SCC cases in the oral cavity, were obtained. All samples (formalin-fixed, paraffin embedded) were examined immunohistochemically for overexpression of p53 protein with monoclonal antibody BP 53-12. As the result, 1. Twenty-two out of 43 leukoplakia cases, and 29 out of 37 oral SCC cases, were positive for p53 protein. 2. p53 protein was overexpressed in premalignant lesions, especially in the cases with moderate and severe epithelial dysplasia. 3. There was a relation between p53 protein expression and pathological features of leukoplakia (epithelial dysplasia), statistically. 4. There was a relation between p53 protein expression and clinical features of leukoplakia, statistically. 5. Malignant transformation during clinical observation was seen in 11 cases. Nine out of 11 cases were positive for p53 even before malignant transformation. Since in cancer-development cases, p53 staining was detected even before malignant transformation of oral leukoplakia to squamous cell carcinoma, it is indicated that p53 accumulation occurred at a early stage of cancer-development. In conclusion, immunohistochemical analysis of p53 protein is suggested to be useful diagnostic procedure for oral leukoplakia, which may develop into oral SCC.     

Cell proliferation and tumour suppressor gene expression in iodine unstained area surrounding oral squamous cell carcinoma

The purpose of this study was to investigate the relationship between epithelial dysplasia unstained with iodine and the expression of proliferating cell nuclear antigen (PCNA) and/or tumour suppressor gene (p53) and the existence of glycogen. Thirty cases of squamous cell carcinomas arising from the buccal mucosa and floor of the mouth were examined. Iodine unstained areas were diagnosed histopathologically as mild, moderate or severe epithelial dysplasia. Normal oral mucosa stained with iodine was used as a control group. There was no histochemical difference in the distribution or ratio of PAS-positive cells between the control and the mild epithelial dysplasia groups, however PAS stained areas of the moderate and the severe dysplasia groups were significantly decreased. Ultrastructurally, glycogen granules were not recognized in the moderate or severe dysplastic epithelia. Immunoreactive ratios of PCNA and p53 in the moderate and severe dysplastic groups were significantly higher than those of the control and the mild dysplasia groups. The positive ratio of PCNA was higher than that of p53, although the immunostaining patterns of PCNA- and p53-positive cells were quite similar. These results suggest that mild dysplastic epithelia that are stained with iodine may be in the category of normal epithelia, whereas both moderate and severe dysplasia that are un-stained with iodine may be suspected of malignant lesions.     

Assessment of p63 expression in oral squamous cell carcinomas and dysplasias

OBJECTIVES: p63, a p53 homologue, may be associated with tumorigenesis in epithelial tissues through its inhibition of p53 transactivation functions. We sought to determine the pattern and levels of p63 expression in oral dysplasias and carcinomas using standard immunohistochemical staining. We also assessed and compared expression of p53 and a cell proliferation marker in these lesions. STUDY DESIGN: This retrospective cross-sectional survey (n=67) included hyperkeratosis (10), mild dysplasia (9), moderate dysplasia (11), severe dysplasia/in situ carcinoma (10), squamous cell carcinoma (SCC) (22 [9 well differentiated, 7 moderately differentiated, 6 poor differentiated]), and normal mucosa (5). Serial sections were stained immunohistochemically with antibodies to p63 (4A4 recognizing all p63 isotypes), p53 (DO-7), and Ki-67 (MIB-1) proteins. In preinvasive lesions, both the percentage of positive cells and staining patterns (negative, basal, suprabasal) were assessed. In oral SCCs, the percentage of positive cells was assessed. Statistical analysis was done using the Tukey-Kramer multiple comparisons test. RESULTS: A suprabasal p63 staining pattern was evident in keratinocyte nuclei in the entire range of noninvasive lesions studied, including normal mucosa. Most nuclei in invasive SCCs stained positive. When all grades of dysplasia were combined, the percent of p63 positive cells was significantly greater than hyperkeratosis (P < .01), and well-differentiated SCC (P < .001). Moderately differentiated SCC had statistically significant more positive cells than well-differentiated SSC (P < .01). Comparison of serial sections showed different p63 staining patterns compared to p53 or Ki-67 staining patterns. CONCLUSIONS: We conclude that p63 is expressed in oral carcinomas and dysplasias, as determined by immunohistochemical staining with a primary antibody to all isotypes. Neither staining pattern nor percentage of stained cells could be used to differentiate the lesions studied. The statistically significant differences found between some groups are not likely to be of diagnostic value. p63 is not coexpressed with p53 expression or Ki-67 suggesting functional independence. When antibodies to the p63 isotypes become available, oral dysplasias and carcinomas should be reassessed.