Mean Labeling Index Research Articles

Evaluation of Cytotoxic T Lymphocytes and Natural Killer Cell Distribution in Oral Squamous Cell Carcinoma and Oral Epithelial Dysplasia: An Immunohistochemical Study.

Background The tumor microenvironment comprises stromal cells, a few immune cells, vascular channels, and an extracellular matrix. The immune cells play a pivotal role in arresting the development of various tumors by identifying and killing the abnormal tumor cells. These immune cells with cytotoxic function include the natural killer (NK) cells and CD8+ T lymphocytes. Human NK cells express the cell surface marker CD57 and can be identified by using monoclonal antibodies. CD8+ cytotoxic T cells are a critical subpopulation of T cells and are important mediators of adaptive immunity. The anti-tumor immunity is important to assess the prognosis of tumors and develop new therapies. This study aimed to evaluate the immunohistochemical expression of CD8 and CD57 immune cells in oral squamous cell carcinoma (OSCC), oral epithelial dysplasia (OED), and normal oral mucosa. Methodology Clinically diagnosed and histopathologically confirmed cases of OSCC (n = 22), oral leukoplakia with OED (n = 22), and normal oral mucosa (n = 22) comprised the study groups. The tissue sections were subjected to immunohistochemical analysis for CD8 and CD57 expression by calculation of the mean labeling index. The results were statistically analyzed using a one-way analysis of variance, Bonferroni multiple comparison test, and Student's t-test. SPSS software version 20.0 (IBM Corp., Armonk, NY, USA) was used for the statistical analysis, and the significance level was set at 0.05. Results An overall statistically significant difference was obtained in the number of CD8+ T lymphocyte cells and CD57+ NK cells when compared between OSCC, OED, and normal oral mucosa (p = 0.01). Variations in the number of CD8+ T lymphocyte cells and CD57+ NK cells were observed when a comparison was made between OED and OSCC and between OSCC and normal mucosal samples (p = 0.01). The study results showed that the mean labeling index of CD8 and CD57 increased in OSCC when compared to OED and normal mucosa (p = 0.01). Conclusions Samples of OED with moderate or severe dysplasia and samples of OSCC were accompanied by a higher level of infiltrating immune cells such as T cells, B cells, NK cells, and macrophages when compared to normal mucosa. The results suggested that the expression of CD8 and CD57 cells increased from normal mucosa to OED and the highest expression was found in OSCC. CD8 and CD57 could be used as surrogate markers to assess the malignant potential of the lesion and to determine the prognosis of patients with oral cancer.

Study of Expression of p63 and Ki-67 in Normal, Dysplastic, and Malignant Lesions of Oral Mucosa

Background: The oral cavity is the most common site of cancer in the Indian subcontinent contributing to one-third of all cancers. The prognosis of oral dysplasia and malignant lesions can be predicted by both clinicopathological parameters and molecular markers. Aims and Objectives: The present study was undertaken to examine and compare the expression profile of cell cycle regulatory proteins p63 and Ki-67 in normal dysplastic and malignant lesions of oral mucosa and its correlation with prognosis. Settings and Design: This was a retrospective case–control study in a tertiary referral center. Materials and Methods: We evaluated and compared the expression profile of antigens p63 and Ki-67 on 60 cases of histologically proven oral dysplastic and malignant lesions oral squamous cell carcinoma (OSCC – 35 cases), oral epithelial dysplasia (OED – 15 cases), and 10 cases of benign and inflammatory oral lesions. Statistical Analysis: Mean and standard deviation were calculated and ANOVA statistical test was used to compare the data. Results: The mean labeling index of p63 and KI-67 was higher in OSCC than OED and minimum in inflammatory (controls) groups. Conclusion: p63 expression had a positive correlation with Ki-67 expression. Thus, p63 and Ki-67 may be a marker for tumorigenesis and tumor cell proliferation.

Characterization of genotypic disorders in ERα+ breast cancers by evaluating ERα and TFF1 expression: Is TFF1 a promising predictive biomarker for endocrine therapy?

TFF1 is overexpressed in estradiol-dependent invasive breast carcinomas Estrogen Receptor positive (ERα+). However, certain subclasses of ERα+ tumors do not overexpress this protein and present a therapeutic escape from endocrine therapy. This study was conducted to characterize the genotypic disorders of ERα+ breast cancers by quantitative evaluation of intratumoral heterogeneity in ERα and TFF1 expression and to precise the clinical significance of TFF1 overexpression in primary ERα+ mammary invasive ductal carcinomas (IDCs). To evaluate the intratumoral distribution of ERα and TFF1, 60 primary ERα+ IDCs were used and stereological analysis were conducted. The labeling index (LI) and the mean labeling index (MLI) for the two markers overexpression were evaluated. The coefficient of variation (COV) was used to estimate spatial dispersion of markers. Stereological analysis showed that the mean ERα and TFF1 labeling indexes (MLI) showed a heterogeneous distribution of markers in each tumor with rates fluctuating between 21.59% ± 10.18 and 46.91% ± 10.61 for ERα and between 18.73% ± 6.32 and 34.39% ± 8.71 for TFF1. The COV showed values fluctuating between 16.75% and 44.40% for ERα and between 25.3% and 65.7% for TFF1 reflecting an important heterogenous dispersion of ERα and TFF1 protein within the same tumor. This study allowed us a simple quantitative estimation of the phenotypic heterogeneity wich is the reflection of genotypic disorders. Our results showed that ERα+ breast cancers are genetically unstable and can present 2 different phenotypes [ERα+/TFF1+] or [ERα+/TFF1¯]. This genetic instability explains the resistance of these cancers to hormonal therapy.

Potentiated action on the progression of OSMF by hypoxia mediated signaling pathway by the epithelial mesenchymal transition and angiogenic apparatus.

Epithelial-mesenchymal transition (EMT) is a complex process, in which epithelial cells acquire the characteristics of invasive mesenchymal cells. EMT has been implicated in cancer progression and metastasis as well as the formation of many tissues and organs during development. The aim of the study was to ascertain the role of hypoxia-mediated signaling pathways influencing EMT and angiogenesis in progression of oral submucous fibrosis (OSMF). Evaluation of the immunoexpression of alpha-smooth muscle actin (α-SMA), E-cadherin, vimentin, and factor VIII receptor antigen in OSMF and oral squamous cell carcinoma (OSCC) arising from OSMF was done. Differences between the different variables were analyzed using ANOVA test and Pearson's Chi-square test, and Mann-Whitney test was also calculated. The mean α-SMA positive myofibroblasts increased from Group 1 (OSMF) to Group 2 (OSCC), especially those in the deeper connective tissue stroma. The mean labeling index of vimentin and mean vessel density immunoexpression was more in Group 2 (OSCC) as compared to Group 1 (OSMF). Mean α-SMA correlated negatively with E-cadherin expression and positively with vimentin and factor VIII immunoexpression. E-cadherin expression correlated negatively with factor VIII and positively with Vimentin expression. The molecular mechanisms responsible for the development of OSCC in patients with OSMF require unification of multiple progressive pathogenetic mechanisms involved in the progression of the disease.

Immunohistochemical expression and evaluation of cyclin D1 and minichromosome maintenance 2 in oral squamous cell carcinoma and verrucous carcinoma

Background:The study of cell proliferation is important for assessing the tumor behavior, prognosis and patient survival of oral carcinomas. As literature search did not reveal sufficient studies of immunohistochemical expression of cyclin D1 and minichromosome maintenance 2 (MCM2) in oral squamous cell carcinoma (OSCC) and verrucous carcinoma (VC), the present study was undertaken.Materials and Methods:The study group included 20 cases of histopathologically diagnosed OSCC, 10 cases of VC and 10 cases of normal mucosa (NM). All samples were evaluated for the expression of cyclin D1 and MCM2 using standard Immunohistochemistry (IHC) procedure.The present study involved both qualitative and quantitative analyses. Qualitative analysis was done by evaluation of intensity and area of staining. Quantitative analysis was done by calculating the percentage of positively stained cells and assessing the labeling index (LI). Data obtained were subjected to statistical analysis using SPSS statistical package (version 23.0).Results:On evaluating and comparing the intensity of staining and area of staining of cyclin D1 and MCM2 between the study groups, statistically significant values (P < 0.05) were obtained using Kruskal–Wallis ANOVA. Comparison of LI of cyclin D1 and MCM2 in NM, OSCC and VC statistically significant results (P < 0.05) was obtained using Mann–Whitney U-test. Mean LI of MCM2 was found to be significantly higher than mean LI of cyclin D1 in all the study groups.Conclusion:From the present study, we conclude that MCM2 has the potential to serve as a novel cell proliferation biomarker in OSCC and VC when compared to cyclin D1.

Intestinal-type gastric dysplasia in Helicobacter pylori-naïve patients.

Gastric dysplasia and gastric cancer in Helicobacter pylori (Hp)-naïve patients usually exhibit a gastric phenotype, reflecting gastric mucosa without intestinal metaplasia (IM). We showed that intestinal-type gastric dysplasia (IGD) rarely occurs in the Hp-naïve stomach. In the last 10years, we treated 1760 gastric dysplasia and gastric cancer patients, with 3.6% (63/1760) being Hp-naïve. Among these, ten were diagnosed with 14 IGDs and enrolled in this retrospective analysis. All lesions were observed by white-light endoscopy (WLE) and narrow-band imaging with magnification endoscopy (NBIME). We analyzed their endoscopic and microscopic features and patient demographics. Five men and five women aged 64 ± 21years were included. WLE showed the depressed lesions mimicking a benign raised erosion in the prepyloric compartment. Multiple growths were confirmed in 30% (3/10) of patients. NBIME showed a near-regular microstructure and capillaries in 50% (7/14) of lesions with a gastritis-like appearance. Histologically, background mucosa was non-atrophic pyloric gland tissue, but 40.0% of samples (4/10) contained sporadic IM. Most of the lesions (8/14) were low-grade dysplasia, and others had a high-grade component, with one progressing to intramucosal carcinoma. The neoplastic surface was widely covered with foveolar epithelium in 57.1% (8/14). Immunohistochemically, neoplastic cells expressed CDX2 in all patients (14/14), MUC2 and CD10 in 92.9% (13/14), MUC5AC in 14% (2/14), and no expression of MUC6, showing an intestinal phenotype. Ki-67 was overexpressed with a mean labeling index of 58.3 ± 38.5%, and p-53 was overexpressed in 92.9% (13/14), regardless of the dysplastic grade. The IGD rarely occurs in Hp-naïve patients with distinctive clinicopathologic characteristics.

Sporadic foveolar-type gastric adenoma with a raspberry-like appearance in Helicobacter pylori-naïve patients.

Sporadic foveolar-type gastric adenoma (FGA) has been described as an extremely rare polyp that is whitish and flatly elevated. However, we recently found that sporadic FGA with a raspberry-like appearance (FGA-RA) is not rare in Helicobacter pylori (H. pylori)-naïve gastric mucosa. We endoscopically or surgically treated 647 patients with gastric epithelial neoplasms in the last 5 years, with 7.7% (50/647) being H. pylori-naïve. Among these, 43 FGA-RAs were diagnosed based on histologic and endoscopic features in 34 patients, who were all enrolled in this retrospective study. All lesions were observed by white-light endoscopy (WLE) and narrow-band imaging with magnification endoscopy (NBIME). We subsequently analyzed their endoscopic and microscopic features and patient characteristics. The patients were 22 males and 12 females aged 57±23 years (mean±2SD). WLE showed raspberry-like small polyps mimicking gastric hyperplastic polyps in the oxyntic gastric compartment (body/fundus). Multiple growths were confirmed in 20.6% (7/34) of the patients. NBIME revealed irregularly shaped papillary/gyrus-like microstructures with abnormal capillaries. Histologically, all lesions were intraepithelial neoplasms, and most of lesions (62.8%, 27/43) exhibited low-grade dysplasia. Immunohistochemically, neoplastic cells featured strong and diffuse MUC5AC expression, negative or very low MUC6 expression, and negative MUC2/CD10 expression. They also showed Ki-67 hyperexpression with a mean labeling index of 59.4±48.7%. The coexistence of fundic gland polyps in the background mucosa was significantly higher in multiple FGA-RA cases than in solitary cases (100% vs. 55.5%, P< 0.05). FGA-RA is a newly suggested histologic variant of sporadic FGA whose occurrence is not rare in daily endoscopic practice.

Intra-tumoral distribution of Ki-67 and Cyclin D1 in ER+ mammary carcinoma: quantitative evaluation.

In spite of the strong evidence demonstrating the role of overexpression of Ki-67 and Cyclin D1 markers in breast carcinomas, clinical and pathological data remain to be discussed. This can be explained partly by intratumor heterogeneity. To define the prevalence and clinical significance of Ki-67 and Cyclin D1 overexpression in primary breast tumors ER positive, while highlighting the existence of intratumor heterogeneity in this type of cancer. 51 ER positive breast cancer tumors were used to evaluate the intratumoral distribution of Ki-67 and Cyclin D1 expression. Image acquisition and visualization of the markers were performed by optical microscopy and stereology sampling method. The mean Ki-67 labeling index was distributed heterogeneously in the same tumor, from 20.67±6.87 to 45.10±10.65. The coefficient of variation (COV) revealed dispersion values between 13.4% and 42.9%. Associated with positive ER status, all the tumors presented a Cyclin D1 expression with a COV varying between 19% and 28.5% and a mean labeling index fluctuating between 19.40±4.42 and 41.64±10.08 within the same patient showing important intratumor heterogeneous distribution. In this study, we have adopted a strictly quantitative approach to evaluate and demonstrate intratumor heterogeneity. This establishes one of the main factors for poor response to cancer therapy. To achieve this, intratumor heterogeneity should be usually definable and quantifiable but this domain awaits future progress and methods need to move towards a better understanding of molecular and cellular mechanisms that initiate and maintain this tumor heterogeneity.

MCM3 proliferative index is worthier over Ki-67 in the characterization of salivary gland tumors.

Salivary gland tumors bear uncanny characteristics of being different based on their morphological aspects rather than the presence of clear demarcation. This ambiguity in the spectrum from benign to malignant salivary gland neoplasms while categorizing the neoplasm is having inherent pitfalls. The present study was, therefore, designed to characterize benign and malignant salivary gland tumors based on their proliferative indices. Study samples comprised of 97 cases of histopathologically confirmed benign and malignant salivary gland tumors. The cases were immunohistochemically assessed for MCM3 and Ki-67 expressions and the molecular characterization was performed based on the findings. The majority of benign and malignant salivary gland tumors were from the parotid gland, (51.2%) and (42.4%), respectively. Overall mean labeling index of MCM3 was higher i.e., (5.60 ± 3.99) in comparison to Ki-67 i.e., (2.82 ± 3.14) with P = 0.05 using paired t-test. Besides, malignant salivary gland neoplasms represented a higher mean score of MCM3 and Ki-67 than benign neoplasms. The requirement of a novel marker has led to the use of MCM3 which has a characteristic role in the entire spectrum of the cell cycle. The present study highlighted the extrapolation of MCM3 over Ki-67 for diagnosis and for true characterization of biologic behavior of salivary gland pathologies which may, in turn, influence the treatment modality employed for such lesions.

Correlation of p63 protein expression and Ki-67 labelling index with histological grades of meningiomas: A five years study

Introduction: Meningiomas account for 13%–25% of primary intracranial tumours and ~25% of intraspinal tumors and histological grading has prognostic and therapeutic implications. However, histological criteria are sometimes not fulfilled to predict the biological behaviour of meningiomas, necessitating use of adjunct ancillary techniques like immunohistochemical markers. The aim of the study was to assess the correlation between the p63 protein expression with MIB-1 labelling index in different histological grades. Materials and Methods: This was a retrospective and prospective study of patients with intracranial and intraspinal meningiomas diagnosed during the period 2009-2014. The H& E Stained histological sections of the entire meningioma tumour were graded according to WHO 2007 criteria. Immunohistochemistry was performed using p63 protein and Ki-67. Result: The age of the patients ranged from 4-75 years including seven children. Out of 135 cases, 46 (34.07) cases occurred in males and the remaining 89(65.92%) cases were females. There were 125 grade I, 06 grade II, 04 grade III meningiomas. Amongst Grade I tumours (n =125) 16% (n=20) exhibited nuclear positivity for p63 protein. Among Grade II (n= 06) and Grade III (n= 04), all showed strong immune expression for p63. Statistical significant increase in mean labelling index (MIB1) was noted between Grade I to II and Grade II to III. Conclusion: Increased p63 expression and Ki-67 labelling index is associated more with high grade Meningioma and possible recurrence. Keywords: Meningiomas, MIB1 labelling index, p63 protein.

Differential Expression of EZH2 and H3K27me3 in Oral Verrucous Carcinoma and Oral Verrucous Hyperplasia.

Enhancer of zeste homolog 2 (EZH2), a component of the polycomb repressive complex 2 that catalyzes trimethylation of H3K27 (H3K27me3), has been shown to promote tumor development and progression. Expression of EZH2 is associated with cell cycle regulation and cell proliferation in various neoplasms. Oral verrucous hyperplasia (OVH) and Oral verrucous carcinoma (OVC) are rare entities and share several clinical and histopathologic features. Problems distinguishing these lesions are added by a lack of adjacent normal tissue of the biopsy samples and poorly oriented tissue sections. The aim of this study was to investigate the expression of EZH2 and H3K27me3 in OVH and OVC and comparing the expression with normal oral mucosa and oral squamous cell carcinoma (OSCC). Seventy-eight samples, including 25 cases of OVC, 8 cases of OVH, 35 cases of OSCC and 10 cases of normal oral mucosa, were retrieved and submitted for immunohistochemical staining. The results demonstrated that the mean labeling indices (LIs) of EZH2 and H3K27me3 expression were highest in OSCC, followed by the OVC, OVH, and normal mucosa. Statistical differences in EZH2 LI were observed among these lesions whereas H3K27me3 LI was significantly different among OSCC, OVH and normal mucosa. EZH2 LI was found to have a sensitivity of 72.00% and specificity of 87.50% in distinguishing OVH from OVC, and a sensitivity of 57.14% and specificity of 84.00% in distinguishing OVC from OSCC. A positive correlation between EZH2 and H3K27me3 expression was significantly found in OVC but not in OVH and OSCC. These findings highlight the involvement of epigenetic regulation by EZH2-mediated H3K27me3 in the pathogenesis of OVH and OVC, and EZH2 expression indicates disease progression of these verrucous lesions. Diagnostic test analysis further suggests that EZH2 may be used as an additional test for differentiating OVH from OVC in questionable cases.

Immunohistochemical profile of the anti-apoptosis, apoptosis and proliferation markers Bcl-2, caspase-3, p53, and Ki-67 in botryoid odontogenic cysts compared to lateral periodontal cysts and gingival cysts of the adult

ABSTRACT We investigated the differences in growth and rates of recurrence of the botryoid odontogenic cyst (BOC) and the less aggressive lateral periodontal cyst (LPC) and gingival cyst of the adult (GCA). We compared the immunohistochemical expression of selected biomarkers of apoptosis and proliferation and of regulators of their activity. Sections from archival paraffin blocks of 15 BOCs, six GCAs, six LPCs, and three odontogenic keratocysts (OKCs) were processed for immunohistochemical localization of Bcl-2, caspase-3, p53 and Ki-67. Labeled and unlabeled epithelial cells were counted and differences in the mean labeling index (LI) were evaluated statistically. The only significant differences in LI were for the anti-apoptotic marker, Bcl-2; the hierarchy was BOC > OKC > LPC > GCA. In two BOCs, 97% of the cells, and in all OKCs, all of the basal cells were labeled with Bcl-2. Otherwise, cells labeled with Bcl-2, p53 and caspase-3 were scattered among the basal and intermediate epithelial cell layers. Ki-67 labeled almost exclusively basal cells in the BOCs, LPCs and GCAs, and both basal and intermediate layer cells in the OKCs. Our findings suggest that while there was no significant difference in replicative potential of the GCAs, LPCs and BOCs, factors that influence apoptosis may be partially responsible for the more aggressive behavior of BOCs and OKCs.

Study of Hypoxia-inducible factor-2α expression in the malignant transformation of Oral submucous fibrosis.

Context:Hypoxia-inducible factor (HIF)-2α is overexpressed in primary and metastatic human cancers, whose expression is correlated with tumor angiogenesis and patient mortality. HIF plays a role in the progression of fibrosis in oral submucous fibrosis (OSF).Aim and Objective:The aim is to study and compare the expression of HIF-2α in OSF (a), oral squamous cell carcinoma (OSCC) with areca nut usage (b), OSCC without areca nut usage (c) and normal mucosa (d) by immunohistochemistry.Subjects and Methods:Immunohistochemical detection of HIF-2α was done on 51 samples, which included 11 cases (a), 15 cases (b), 15 cases (c) and the expression was compared with that of (d).Statistical AnalysisData were analyzed using the SPSS™ software (ver. 21.0). Chi-square test and kappa analysis were performed to compare the intensity of staining between the groups and for inter-observer agreement, respectively. Value of P ≤ 0.05 was considered statistically significant. The mean labeling index between the groups was studied by the Kruskal–Wallis test.Results:All the cases of (d), (a), (b) and (c) showed HIF-2α expression (P = 0.329). About 13% cases of (c) showed intense expression (P = 0.406) and 27% of (a) cases showed expression only in the connective tissue (P = 0.023). The number of positively stained nuclei in both (b and c) cases reduced as the tumor progression was from well to poorly differentiated.Conclusion:Areca nut initiates fibrosis and subsequent hypoxia in OSF which triggers HIF-2α expression in the epithelium. HIF-2α could be a surrogate marker for cancer initiation and progression.

Ki67 labeling in CNS tumours

A study of Ki67 Labeling Index (LI) by immunohistochemistry was carried out in 47 CNS neoplasms. Astrocytic and oligodendroglial tumours were the most common group with glioblastoma, NOS and diffuse astrocytoma, NOS Grade II predominating. Ki67 LI of diffuse astrocytomas varied from 0.72 to 13.9% with a mean of 4.36%. In glioblastomas the Ki67 LIs varied from 5.96 to 22.4% with a mean of 12.3% which was comparable with various studies. 1 pleomorphic xanthoastrocytoma was reclassified as anaplastic type based on a high LI of 21.66%. All histological variants of ependymomas showed uniformly low Ki67 LIs. Grade I meningiomas show ed a mean Ki67 LI of 2.22% for benign, 5.85% for atypical Grade II and 30% in the anaplastic Grade III meningioma. Choroid plexus carcinoma and Non Hodgkins lymphomas showed very Ki67 LIs from 60 to 91%. The findings correlated well with WHO CNS tumour grade and helped to identify histologically defined lower grade tumours with potential for recurrence and progression. Keywords: CNS neoplasms, Gliomas, Proliferation markers, Ki67.

MCM 2 a novel marker in predicting recurrence and prognosis at negative surgical margins of oral squamous cell carcinoma

ObjectivesThe treatment failure in Oral carcinoma patients is mainly due to recurrence leading to a poor prognosis. Genetically transformed cells in the adjacent mucosal area thought to be the reason for local recurrences and an invasive tumour front area is known to host the aggressive tumour cells. Hence, the study conducted to quantify and compare proliferative markers Ki-67 and MCM 2 antigens at the margins and invasive tumour front to predict recurrence and prognosis in Oral squamous cell carcinomas. Materials and methodThe study involved paraffin tissue sections of 30 cases of recurrent, and 30 cases of non recurrent Oral squamous cell carcinomas were subjected to Immunohistochemical analysis at negative surgical margins and invasive tumour front (ITF). The mean labelling index (Li) of MCM 2 (Minichromosome Maintenance Protein 2) and Ki-67 was compared among the groups with ‘t’ test to predict the recurrence and overall survival by Kaplan-meier curve and Log-rank test survival estimate. ResultsThe Li of Ki-67 and MCM 2 were higher at negative margin and ITF of recurrent group compared to non recurrent OSCC group with statistically significant difference (P < 0.05) only with MCM 2. Li of MCM 2 at margin and ITF was a better predictor of overall survival than Ki-67. The overall survival was significantly lower in 43.95 months with Li of MCM 2 more than 48.2 at margin. ConclusionMCM 2 is a novel marker at negative margins in predicting the recurrence and survival of Oral squamous cell carcinoma.

Immunohistochemical identification of salivary pleomorphic adenoma subtypes using p63 and alpha smooth muscle actin myoepithelial markers

Background: Pleomorphic adenoma (PA) histopathologically represents a heterogeneous lesion with varying proportion of mesenchymal and epithelial tissue. Due to its heterogeneity, identifying various subtypes often presents a diagnostic dilemma. Hence, the aim of this study is to determine and compare the activities of salivary myoepithelial (myo) and basal reserve cells (BRC) within the subtypes of salivary PA using p63 and alpha smooth muscle actin (α-SMA) immunohistochemical markers. Method: In this retrospective study, 24 archival tissue blocks of PA (12 stroma-rich, 6 cell-rich, and 6 classic subtypes) retrieved and subjected to immunohistochemistry using monoclonal p63 and α-SMA antibodies. Fraction of positive cells expressed as the percentage of total cells within each sample, labeling indices (LI) was computed. Result: PA samples subjected to immunohistochemistry showed 100% p63 and 95.9% α-SMA immunoreactivity. Statistically significant difference (p = 0.01) was observed with p63 mean LI across PA subtypes. Overall, p63 LI (37.1 ± 13.5) was higher than α-SMA LI (26.5 ± 9.5). Staining pattern for both markers was similar within epithelial and stroma elements. Conclusion: Disparity in Mean LI of the markers with higher index observed for p63, suggests additional activity of BRC, whose variable expression is associated with the differential ratio of epithelial to stroma components within PA subtypes.

Chromosomal aberrations in chordoid meningioma - An analysis.

Chordoid meningiomas (CMs) are a rare subgroup of tumors, accounting for approximately 0.5% of all meningiomas. These tumors correspond to World Health Organization (WHO) Grade II lesions and behave aggressively, with an increased likelihood of recurrence. There are only two studies that have described the genetic alterations in CMs. While a majority of meningiomas are known to have deletion at many chromosomal loci such as 22q, 18p, 14q, and 1p, which are found to be associated with initiation, progression, and malignancy of these tumors, these have not yet been studied in CMs. Thus, our aim was to evaluate the status of these four chromosomal aberrations in CMs and correlate the findings with the clinical outcome of patients. A total of 15 cases of CM operated over a period of 12 years from 2001 to 2013 were analyzed. The archival paraffin blocks were retrieved and sections were subjected to locus-specific fluorescent in situ hybridization (FISH) using 22q12.2, 18p11.3, 14q32.2, and 1p32.3 probes. Immunohistochemistry (IHC) was done on all cases using MIB-1, vimentin, glial fibrillary acidic protein (GFAP), and epithelial membrane antigen (EMA) antibodies. All cases had characteristic features of CM, and were positive for EMA and vimentin and negative for GFAP. The mean labeling index for MIB-1 was 2.7 ± 0.8%. Of the 15 cases, 5 cases showed recurrence with a median follow-up period of 28 months. Patients who underwent Simpson's grade I excision did not show any relapse of the tumor. Of the 5 recurrent cases, 4 had complete deletion of all four chromosomal loci. Among the 10 nonrecurrent cases, 9 (90%) showed either partial deletion or an intact status. This is the first study to evaluate the combined chromosomal status of 22q, 18p, 14q, and 1p in CMs. Our study shows that there was a higher propensity of recurrence in tumors, even with complete excision, with complete deletion in all four chromosomal loci.

CKS1 expression in melanocytic nevi and melanoma.

Cyclin-dependent kinase subunit 1 (Cks1) regulates the degradation of p27, an important G1-S inhibitor, which is up regulated by MAPK pathway activation. In this study, we sought to determine whether Cks1 expression is increased in melanocytic tumors and correlates with outcome and/or other clinicopathologic prognostic markers. Cks1 expression was assessed by immunohistochemistry in 298 melanocytic lesions. The frequency and intensity of cytoplasmic and nuclear expression was scored as a labeling index and correlated with clinico-pathological data. Nuclear Cks1 protein was found in 63% of melanocytic nevi, 89% primary and 90% metastatic melanomas with mean labeling index of 7 ± 16, 19 ± 20, and 30 ± 29, respectively. While cytoplasmic Cks1 was found in 41% of melanocytic nevi, 84% primary and 95% metastatic melanomas with mean labeling index of 18 ± 34, 35 ± 34, and 52 ± 34, accordingly. Histologic stepwise model of tumor progression, defined as progression from benign nevi to primary melanomas, to melanoma metastases, revealed a significant increase in nuclear and cytoplasmic Cks1 expression with tumor progression. Nuclear and cytoplasmic Cks1 expression correlated with the presence of ulceration, increased mitotic rate, Breslow depth, Clark level, tumor infiltrating lymphocytes and gender. However, other well-known prognostic factors (age, anatomic site, and regression) did not correlate with any type of Cks1 expression. Similarly, increasing nuclear expression of Cks1 significantly correlated with worse overall survival. Thus, Cks1 expression appears to play a role in the progression of melanoma, where high levels of expression are associated with poor outcome. Cytoplasmic expression of Cks1 might represent high turnover of protein via the ubiquination/proteosome pathway.

Botryoid odontogenic cyst. Exploration of proliferative activity, apoptosis and expression of TP53 and BCL2 compared to the histologically identical lateral periodontal and gingival cysts

The botryoid odontogenic cyst (BOC) is a rare, locally more aggressive variant of the usually indolent lateral periodontal cyst (LPC) and gingival cyst (GC). A recent case of BOC provided an opportunity for an exploratory study on the causes of its more aggressive behavior. The limited objective was to see if the BOC was sufficiently different from the other cysts to warrant an investment in a large study. Sections of neutral buffered formalin fixed, paraffin-embedded tissues from the BOC and archival specimens of four GCs, four LPCs and three odontogenic keratocysts (OKCs) were stained using immunohistochemistry for Ki-67, a marker of proliferating cells, caspase-3, a marker of cells undergoing apoptosis, tumor suppressor p53, and the apoptosis inhibitor BCL2. The mean labeling index (LI) of immunoreactive cyst epithelial cells was computed for each antibody and type of cyst. Compared to the LPCs and GCs, the BOC exhibited a moderately larger Ki-67/caspase-3 LI difference, which indicates that the BOC had a net higher rate of growth. We found a much higher level of LI, therefore likely dysregulation of p53. We also found a much higher LI of BCL2. The LIs of p53 and BCL2 in the BOC were similar to and more than twice that of the OKCs, respectively. Although meaningful statistical analysis was precluded by our use of only one case of BOC and a small number of the other cysts, the high p53 and very high BCL2 labeling indices of the BOC offer a potential explanation for its reportedly more aggressive behavior that clearly is worthy of further investigation.

Increased NG2 and SOX2 expression is associated with high-grade choroid plexus tumors.

The World Health Organization classification of choroid plexus tumors (CPT) includes three distinct grades: Choroid plexus papilloma (CPP), atypical choroid plexus papilloma (ACPP) and choroid plexus carcinoma (CPC). The molecular basis for these pathological distinctions may help to stratify tumors and provide an insight into the clinical behavior of CPTs. In the present study, the progenitor and stem cell markers neuron glia antigen-2 (NG2) and sex-determining region Y-box 2 (SOX2) were investigated as potential biomarkers that may distinguish between distinct CPT grades. Immunohistochemistry was used to determine the expression of NG2 and SOX2 in CPTs (n=34) from Chinese patients (21 males and 13 females) with a mean age of 31.1 years (range, 1-63 years). The proportion of cells stained were scored using a scale between 0 and 3+, where 0 represents no staining and 3+ represents strong staining, and mean scores for each marker were determined on the basis of tumor grade. Pathological diagnosis revealed a distribution of cases as follows: CPP, 25; ACPP, 5; and CPC, 4. NG2 and SOX2 were expressed in CPTs of all grades. The mean labeling indices for CPP, ACPP and CPC were 1.12, 1.80 and 2.75 for NG2, respectively, and 1.20, 2.00 and 3.00 for SOX2, respectively. Statistical analysis of the mean labeling indices revealed a significant association between the expression of NG2 and SOX2 and CPT grade (P=0.001 and <0.001 for CPP/ACPP and CPP/CPC, respectively). The results of the present study indicated that increased expression of NG2 and SOX2 was associated with higher-grade tumors and that these markers may be useful in determining CPT grade.