Cyclin D1 and Ki-67 expression correlates to tumor staging in tongue squamous cell carcinoma

Background

The immunohistochemical expression of Cyclin D1 and Ki-67 were analyzed in tongue squamous cell carcinomas (SCC), relating them to the clinical and morphological exhibition of these tumors.

Material and Methods

Twenty-nine patients fulfilled the inclusion criteria; clinical data included gender, age, ethnicity and use of licit drugs such as alcohol and tobacco. The TNM staging and histopathological differentiation grading was assessed for each case. In addition, T1 patients were gathered with T2 patients; and T3 patients were gathered with T4 patients to assemble two distinct groups: (T1/T2) and (T3/T4).

Results

The mean follow-up time was 24 months and 30% of the patients died as a consequence of the disease, while 23.3% lived with the disease and 46.7% lived lesion-free. T1 and T2 tumors showed statistically lesser Ki-67 and Cyclin D1 staining when compared to T3 and T4 tumors.

Conclusions

Ki-67 and Cyclin D1 pose as auxiliary tools when determining the progression of tongue SCC at the time of diagnosis. Key words:Carcinoma, squamous cell, cyclin D, immunohistochemistry, Ki-67 antigen, prognosis.     

PCNA and Ki-67 immunoreactivity in multinucleated cells of giant cell fibroma and peripheral giant cell granuloma

Immunohistochemical investigation of PCNA and Ki-67, two diverse nuclear proteins essential to the cell cycle, was undertaken in archival, formalin-fixed and paraffin-embedded specimens of giant cell fibroma (GCF) and peripheral giant cell granuloma (PGCG). GCF multinucleated cell nuclei were mostly PCNA⁺, although there was variability in staining intensity. This indicates heterogeneity in nuclear PCNA metabolism of GCF multinucleated cells, and it is possible that the most intensely stained nuclei have passed through the cell cycle more recently compared to the less immunoreactive nuclei. However, the absence of Ki-67 immunoreactivity in GCF multinucleated cells, and absence of mitoses in GCF multinucleated cells, suggests that cell cycling in the absence of cytokinesis is not involved in GCF multinucleated cell formation. Alternatively, GCF multinucleated cells possibly form by fusion of mononuclear cells previously identified as fibroblasts, although this theory cannot be confirmed by the data presented in this study, and the histogenesis of GCF multinucleated cells remains unclear. In contrast, absence of either PCNA or Ki-67 immunoreactivity in PGCG multinucleated cells is consistent with an osteoclast lineage and formation from differentiated mononuclear cells.     

PCNA/AgNOR and Ki-67/ AgNOR double staining in oral squamous cell carcinoma

This study was performed on oral squamous cell carcinomas (OSCC) in order to investigate the relation between the number of interphase silver-stained nucleolar organizer regions (AgNORs) and the immunolabeling of proliferation-associated markers, using antibodies to Ki-67 and proliferating cell nuclear antigen (PCNA). Fifteen consecutive cases of oral squamous cell carcinoma were used and a double staining technique was performed in order to quantify the number of NORs in PCNA-positive and -negative cells as well as in Ki-67-positive and -negative cells. Our results showed a higher mean number of AgNORs in PCNA- and Ki-67-positive cells than in PCNA- and Ki-67-negative cells. We concluded that there is an association between cell proliferation and AgNOR score in OSCC.     

The value of Phosphohistone H3 as a cell proliferation marker in oral squamous cell carcinoma. A comparative study with Ki-67 and the mitotic activity index

Background The Phosphohistone H3 (PHH3) antibody is recognized as a biomarker of cell proliferation, specific for cells in mitosis, of prognostic value in different malignant neoplasms, however it has been poorly studied in oral squamous cell carcinoma (OSCC). The main objective of this study was to evaluate the immunoexpression of the PHH3 in the OSCC, through the correlation with the immunoexpression of Ki-67, the mitotic activity index (MAI), histological grading, clinical-morphological parameters and the rate of survival.Material and Methods The study sample consisted of 62 cases of OSCC diagnosed in the Pathological Anatomy Laboratory of the Faculty of Dentistry, University of the Republic (Uruguay). In each of them, an immunohistochemical technique was performed for Ki-67 and PHH3 (serine 10) antibodies. Image J software was used for the MAI and biomarker quantification, defining the percentage of positivity and mitotic Figures per 1000 tumor cells.Results a significant association was obtained between the expression of PHH3 (p 0.016) and MAI (p 0.031) with survival time. However, no similar relationship was found with Ki-67 (p 0.295). Although it was confirmed a statistical association between histological grade and Ki-67 immunoexpression (p 0.004), PHH3 did not show a similar relationship (p 0.564).Conclusions It was confirmed the role of the PHH3 antibody as a biomarker of mitotic Figures in OSCC and as a potential marker of cell proliferation. It is noteworthy that this is one of the first works that evaluates a possible relationship between the expression of this antibody and survival in OSCC. Key words:Oral squamous cell carcinoma, phosphohistone H3, Ki-67, cell proliferation.     

PCNA, Ki-67 and p53 expressions in submandibular salivary gland tumours

Salivary gland tumours are uncommon with a broad heterogeneity. The most common benign tumour is the pleomorphic adenoma, whereas mucoepidermoid carcinoma and adenoid cystic carcinoma predominate among the malignancies. Most salivary gland tumours occur in the parotid, and consequently clinical and biological data are normally derived from this site. This work describes the expressions of PCNA, Ki-67 and p53 in 15 pleomorphic adenomas, 15 mucoepidermoid carcinomas and 15 adenoid cystic carcinomas of the submandibular gland. Our results showed that all pleomorphic adenomas were negative for p53 and Ki-67 with 66.6% being positive for PCNA. Conversely, p53 was positive in 53% of the mucoepidermoid carcinomas and in 20% of the adenoid cystic carcinomas. Ki-67 was expressed in 47.7% of the mucoepidermoid carcinomas and 40% of the adenoid cystic carcinomas. All malignant tumours were positive for PCNA. These results indicate that the proliferative rate analysed with PCNA and Ki-67 and the expression of p53 in pleomorphic adenoma and adenoid cystic carcinoma of the submandibular gland were similar to those described in the parotid and minor salivary glands. However, mucoepidermoid carcinomas showed higher expression of these markers than those of other salivary glands. This work is the first describing the expression of these immunohistochemical markers exclusively in submandibular salivary gland tumours.     

Evaluation of p53, Caspase-3, Bcl-2, and Ki-67 markers in oral squamous cell carcinoma and premalignant epithelium in a sample from Alava Province (Spain)

Objectives: The objective of this study was to determine whether alterations in the expression of p53, caspase-3 Bcl-2, and ki-67 appear early in premalignant oral epithelium and show clonal behavior. Study Design: Samples from 41 tumors with their adjacent non-tumor epithelia were immunohistochemically analyzed using monoclonal antibodies that recognize p53, caspase-3, Bcl-2, and Ki-67 Results: A statistically significant association was found between the expression in tumor and adjacent epithelium of p53, caspase-3, and Bcl-2 but not of k-67. A significant association was observed between the expression of ki-67 and p53 in both localizations. In non-tumor (premalignant) epithelium samples, there was a significant inverse relationship between the expressions of p53 and caspase-3 and a significant direct relationship between the expressions of p53 and Bcl-2. Conclusions: Alterations in these proteins appear to operate in combination with premalignant epithelia to create hyperproliferative cell states that favor the acquisition of summative oncogenic errors that confer invasive capacity. Key words:Cell cycle, apoptosis, p53, caspase-3, Bcl-2, Ki-67.     

Immunohistochemical expression of p53, MDM2, Ki-67 and PCNA in central giant cell granuloma and giant cell tumor

Central gaint cell granuloma (CGCG) is a reactive bone lesion that occurs mainly in the jaws. The gaint cell tumour (GCT) is a benign locally aggressive neoplasm located near the articular end of tubular bones. Both lesions are characterised histologically by multinucleated gaint cells in a background of ovoid to spindle-shaped mesenchymal cells. There is a basic question whether both lesions are separate entities or variants of the same disease. The study of cell cycle-associated proteins may give insights into clarifing such question. The expression of these protiens is also important to determine the cell cycle regulation in both tumours. The purpose of this study was to evaluate the immunohistochimical expression of p53, MDM2, Ki-67 and PCNA in CGCG and GCT. The results demonstrated that, despite the lack of p53 immunoreactivity, all the samples showed wide expression of MDM2. The percentage of Ki-67-and PCNA-positive cell in CGCG was statistically higher than that of GCT. Our findings shwo that CGCG has a higher proliferative activity compared with that of the GCT. Our results also suggest that p53 inactivation by MDM2 expression may be involved in the pathogenesis of gaint cell lesions of the jaws and long bones.     

p53, PCNA and Ki-67 expression in oral squamous cell carcinomas: the vagaries of fixation and microwave enhancement of immunocytochemistry

Proliferation markers are widely used as indicators of tumour progression and aggression. Fixation and antigen retrieval methods may enhance the immunocytochemical sensitivity of these markers but may also lead to loss of specificity. As these methods are often used quantitatively, standardisation of internal and external methodology is paramount. This study aimed to compare the effects of alcohol and formalin fixation and of microwaving on the immunocytochemical demonstration of p53, PCNA and Ki-67 in oral squamous cell carcinoma using duplicate tissue blocks from 24 cases. Both qualitative and quantitative differences in antigen expression were revealed. Whilst alcohol fixation alone at least maintained and usually increased the strength of positive staining, microwaving alcohol-fixed sections often gave rise to non-specific staining. p53 staining following microwave enhancement of alcohol-fixed tissue showed a significant incidence of conversion of negative results to positive and of positive staining in unexpected tissue components. Alcohol fixation increased the sensitivity of PCNA detection with a far less dramatic loss of specificity. The results emphasise the need for careful standardisation of immunocytochemical methods, particularly when used quantitatively and for inter-laboratory comparisons.     

Correlation of proliferative markers (Ki-67 and PCNA) with survival and lymph node metastasis in oral squamous cell carcinoma: a clinical and histopathological analysis of 113 patients

The purposes of this study were to examine the correlations between proliferation markers and survival rate in oral squamous cell carcinoma (OSCC) patients, and to evaluate the efficacy of proliferation markers in predicting lymph node metastasis. The patients' age, gender, T score, clinical stage, PCNA and Ki-67 index were analysed. Univariate analysis showed that T score had a significant influence on survival, and stage 4 group had a significantly lower survival rate. Lymph node metastasis was also a significant predictor of survival. Using a cut-off point of 25%, those patients with lower Ki-67 scores had survival advantage over those with higher Ki-67 scores. PCNA did not show any differences in survival with a cut-off point of 50%. Ki-67 and PCNA were significantly higher in the primary tumours associated with lymph node metastasis (pN+) than in those without lymph node metastasis (pN0). Multivariate analysis showed that clinical stage and Ki-67 were independent prognostic factors for survival in OSCC patients. From this result, it can be postulated that the cancer staging based on the TNM stage was a powerful prognostic variable and Ki-67 had a significant effect on the cumulative survival rate.     

Immunohistochemical evaluation of cell proliferation antigen Ki-67 and apoptosis-related proteins Bcl-2 and caspase-3 in oral granular cell tumor

PURPOSE: We sought to evaluate the cell proliferation activity and immunohistochemical expression of proteins that promote or inhibit apoptosis in oral granular cell tumor (GCT). STUDY DESIGN: Immunohistochemistry for Ki-67, a cell proliferation marker; Bcl-2, an anti-apoptotic protein; and caspase-3, a protein implicated in the execution of apoptosis, was performed on tissues from 12 patients with GCT of the tongue. RESULTS: Nuclear immunostaining for Ki-67 was observed only in isolated GCs (less than 2%). All patients exhibited cytoplasmic immunoreactivity for Bcl-2 in the majority of tumor cells. Cytoplasmic staining for caspase-3 was also present in more than 50% of GCs in all tumors. CONCLUSIONS: GCT cells display low proliferation activity, a finding possibly related to their benign behavior. Caspase-3 expression suggests activation of the apoptotic cascade in the GCs, but persistence of the cells in the tissues could be attributed to the expression of Bcl-2 protein, a molecule that functions as a survival factor.     

Evaluation of p53, PCNA, Ki-67, MDM2 and AgNOR in oral peripheral and central giant cell lesions

OBJECTIVES: Peripheral giant cell lesion (PGCL) and central giant cell lesion (CGCL) of the jaws have a distinct clinical behaviour. Whether such biological differences are supported by a distinct pattern of proliferation markers or cell cycle associated proteins expression is not known. Therefore the purpose of the present study was to compare the immunohistochemical expression of p53, MDM2, Ki-67, PCNA and the histochemical expression of argyrophilic nuclear organiser region (AgNOR) on PGCL and CGCL of the jaws. MATERIALS AND METHODS: Paraffin wax blocks of 14 cases of PGCL and 12 cases of CGCL were retrieved. A biotin-streptavidin amplified system was used for identification of the antigens. The AgNOR number was also evaluated. RESULTS: Ki-67 immunoreactivity was greater in the mononuclear cells of PGCL compared to CGCL. PCNA and AgNOR staining were similar in PGCL and CGCL. Prominent MDM2 immunoreactivity was observed in all tissues investigated. By contrast, there was no p53 immunoreactivity. CONCLUSIONS: Although CGCL present a more aggressive clinical behaviour, it has a decreased proliferative activity compared to PGCL. Finally, p53, MDM2, PCNA, Ki-67 immunohistochemical expression and AgNOR histochemical expression do not reflect their distinct biological behaviour.     

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.     

Immunohistochemical detection of DNA topoisomerase type II α and Ki-67 in adenoid cystic carcinoma and pleomorphic adenoma of the salivary gland

Immunohistochemical detection of cell proliferation-associated antigens was investigated in 28 cases of adenoid cystic carcinoma (ACC) and 20 cases of pleomorphic adenoma (PA), using antibodies against DNA topoisomerase type II alpha (topo-II) (Ki-S1) and Ki-67 (MIB-1). The correlation of staining indices with clinicopathological data, histological features and prognosis was also studied. The topo-II value was significantly higher in ACC than in PA (P<0.0001), and highest in the solid growth pattern of ACC. In addition, significant relationships were found between topo-II values and clinical features such as local recurrence, surgical margins, and distant metastases. By log-rank test, the topo-II index was also correlated significantly with patient survival (P<0.01). The values of topo-II index paralleled those of Ki-67 index in ACC, and a correlation coefficient of 0.97 was obtained. Topo-II may be considered an additional marker for estimating the proliferating fraction of cells and for predicting the short-term prognosis for patients with salivary gland tumors.     

Peripheral dentinogenic ghost cell tumor—report of two cases and review of the literature

Oral and Maxillofacial Surgery - Peripheral dentinogenic ghost cell tumor (DGCT) is a rare and non-aggressive benign odontogenic tumor. They usually affect the elderly and are predominantly located...     

增殖细胞核抗原及Ki-67在口腔白斑和鳞癌中的表达及其相关性

目的：比较研究增殖细胞核抗原（PCNA）和Ki-67在口腔癌前病变中的表达，方法：选取人正常口腔粘膜（NOM）（8例），口腔白斑（LK）（31例）及口腔鳞状细胞癌（OSCC）（22例）标本，同时和LSAB法染色PCNA和Ki-67表达，分析增殖活性，用SPSS9．0统计软件分析各组间各指标的差异，PCNA指数和Ki-67指数的相关性。结果：在NOM上皮及单纯增生上皮内，PCNA阳性细胞散在分布于上皮基底层细胞，统计学分析二者无差异。在异常增生上皮各组中，随着增生程度的病理分级的增加，PCNA阳性细胞数增加，分布从其底层向表层增加。且不同增生程度各组间统计学上有显著性差异。在高分化鳞癌中，主要分布在癌巢周围及肿瘤浸润前沿区域。Ki-67与PCNA分布特征类似，但其指数普遍低于PCNA指数。Pearson相关系数r=0．79,二者呈线性正相关。结论：PCNA及Ki-67与口腔粘膜增殖异常有关，其增殖指数与增生程度平行，且二者本身呈线性相关。     

Evaluation of pretreatment serum interleukin-6 and tumour necrosis factor alpha as a potential biomarker for recurrence in patients with oral squamous cell carcinoma

Background

Oral squamous cell carcinoma (OSCC) constitutes 3 percent of all cancers with predominant occurrence in middle aged and elderly males. Tumour recurrence worsens disease prognosis and decreases quality of life in patients with OSCC. Proinflammatory cytokines such as interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-α) have been suggested to play a certain role in variety of tumours. The aim of this study was to investigate the relationship of pretreatment serum IL-6 and TNF-α levels on tumour recurrence in patients with OSCC in order to identify potential biomarkers for the early detection of disease recurrence.

Material and Methods

The patients with newly diagnosed OSCC were treated and followed from the first visit from November 2006 until January 2008. Serum IL-6 and TNF-α concentrations were measured. The records of the patients were re-examined in July 2012 and data were recorded about cancer characteristics and tumour recurrence. Disease free survival was analyzed by Kaplan-Meier survival curves, log rank test and Cox proportional hazards regression.

Results

Serum IL-6 was shown as an independent risk factor for tumour recurrence.

Conclusions

Pretreatment serum IL-6 concentration may be a useful biomarker for identification of OSCC patients with increased risk of the disease recurrence. Key words: Serum IL-6, serum TNF-α, oral cancer, recurrence.     

Cellular proliferation markers in peripheral and central fibromas: a comparative study

Objective:

To perform a comparative study of the cellular proliferation in the peripheral and central fibromas.

Material and Methods:

Immunohistochemistry for PCNA and the AgNOR technique were performed in 9 cases of peripheral odontogenic fibroma (POF), in 4 cases of odontogenic fibroma (OdF), in 8 cases of peripheral ossifying fibroma (PEOF) and 7 cases of ossifying fibroma (OsF). The Kruskal-Wallis and Mann-Whitney tests were used for the statistical analyses.

Results:

Mesenchymal component of the central lesions presented a higher mean number of AgNOR per nucleus and PCNA index than did the peripheral lesions (P≤0.05). The mean number of AgNOR per nucleus in the epithelial component proved to be higher in the OdF than in the POF (P≤0.05). The mesenchymal and epithelial components presented similar mean numbers of AgNOR per nucleus and PCNA index in the OdF, as well as a similar mean number of AgNOR per nucleus in the POF.

Conclusions:

The mesenchymal component may well play a role in the differences between the biological behaviour of the central lesions as compared to the peripheral lesions. Moreover, considering that the epithelial and mesenchymal components in odontogenic fibromas presented a similar proliferation index, more research is warranted to understand the true role of the epithelial components, which are believed to be inactive in nature, as well as in the development and biological behaviour of these lesions.     

环氧合酶-2、Ki-67在涎腺基底细胞腺瘤和基底细胞腺癌中的表达及临床意义

目的:研究环氧合酶-2(Cox-2)及Ki-67在涎腺正常组织(normal salivary gland,NSG)、基底细胞腺瘤(basal celladenoma,BCA)和基底细胞腺癌(basal cell adenocarcinoma,BCC)中的表达及相关性,探讨其表达程度及在恶变中的相互关系。方法:采用免疫组化(PV6001二步法)检测NSG 11例、BCA18例、BCC 9例中Cox-2及Ki-67的表达情况;采用SPSS 12.0软件对实验数据进行分析。结果:Cox-2在NSG中表达与BCA、BCC的差异均有统计学意义(P<0.01);Ki-67在NSG与BCA表达差异无统计学意义(P>0.05),在NSG与BCC中的表达差异有统计学意义(P<0.01);Cox-2、Ki-67表达与BCA组织病理分型的差异无统计学意义(P>0.05);Cox-2在不同部位的BCA中表达差异有统计学意义(P<0.01);Ki-67在不同部位BCA中表达差异无统计学意义(P>0.05);Cox-2和Ki-67在BCA表达中呈负相关(P>0.05),rs=-0.21,而在BCC表达中呈正相关,rs=0.94,P<0.01。结论:Cox-2、Ki-67可能参与BCA、BCC的发生、发展过程;联合检测Cox-2和Ki-67有助于BCA、BCC的预后判断。     

Spatial and temporal distribution of Ki-67 proliferation marker, Bcl-2 and Bax proteins in the developing human tooth

Objective

To investigate the spatial and temporal expression of proliferation Ki-67 marker, pro-apoptotic Bax and anti-apoptotic Bcl-2 proteins during early development of the human tooth.

Materials and methods

Histological sections of eight human conceptuses, 5–10 postovulatory weeks old, were used for immunolocalization for Ki-67, Bax and Bcl-2 markers. Quantification was performed by calculating the fraction of Ki-67 positive cells, expressed as a mean ± SD, and analysed by Mann–Whitney test, Kruskal–Wallis and Dunn's post hoc test.

Results

In 6th–7th developmental weeks, the tooth germ and dental crest contained 37% of proliferating cells, which increased to 40% in the 8th week, and then decreased to 15% in the 10th week, whilst the proliferation in the ectomesenchyme subsequently dropped from 37% to 23%. Epithelial parts of the enamel organ displayed similar proliferation activity (31–36%), dental crest 10%, whilst enamel knot showed no proliferating activity. The tooth ectomesenchyme contained more proliferating cells (50%) than the jaw ectomesenchyme (35%), and both dropped to 28% in the 10th week. Ectomesenchyme between the tooth germs contained 23%, whilst the jaw ectomesenchyme contained 15% of proliferating cells. Bcl-2 expression had following pattern: strong in proliferating cells, moderate in tooth germs and dental crest, and weak in the ectomesenchyme. Bax co-expressed with Bcl-2 in the tooth germ and dental crest. In the reticulum and inner enamel epithelium Bcl-2 had prevalent expression, whilst Bax prevailed in the outer enamel epithelium and tooth ectomesenchyme.

Conclusions

Proliferating cells most likely influence growth of the tooth germ, Bcl-2 affects proliferation and differentiation of specific cell lineages, whilst Bax influences process of cell death.     

A retrospective radiographic evaluation of the anterior loop of the mental nerve: Comparison between panoramic radiography and cone beam computerized tomography

Objectives: To compare the prevalence and the length of mental loop, measured with panoramic radiography (PR) and cone beam computerized tomography (CBCT). Material and Methods: PG and CBCT images where analyzed by a single calibrated examiner to determine the presence and the position of the mental foramen (MF), its distance to the lower mandible border, the anterior length of the mental loop (ML) and the bone quality in 82 PR and 82 CBCT. Results: ML was identified in 36.6 % of PR and 48.8 % of CBCT. PR showed a magnification of 1.87 when compared to CBCT. The mean of anterior extension of the inferior alveolar nerve and the distance to the inferior border of the mandible was higher for PR (2.8 mm, sd 0.91 mm on the PR , range 1.5 to 4.7 mm and 1.59, sd 0.9 on the CBCT ,range 0.4 to 4.0 mm) Conclusions: There is a magnification in PR images with respect to those of CBCT. The differences between CBCT and PR with regards to the identification and length of the ML are not statistically significant. Identification and accuracy measurements of ML did not depend on the bone quality. Considering that two dimensional imaging provides less accurate and reliable information regarding the anterior loop, a CBCT scan could be recommended when planning implant placement in the anterior region. Key words:Mental loop, mental nerve, mental canal, preoperative implant planning, panoramic tomography, cone beam computerized tomography.