CD133 Role in Oral Carcinogenesis.

OBJECTIVE: to investigate CD133 immunoexpression, cancer stem cells marker, in oral epithelial dysplasias (OEDs) and oral squamous cells carcinomas (OSCCs) and understandits possible involvement in the malignant transformation process of these lesions and to better elucidate their biological behavior. MATERIAL AND METHODS: Tissue samples of 15 cases of OSCCs and 15 OEDs were subjected to CD133 antibody immunohistochemistry reactions. The analysis used quantitative parameters (number of immunostained cells regardless of immunostaining sublocations). RESULTS: All samples of OSCCs and OEDs showed positive immunostaining, with no significant difference between these groups (p = 0.283). We did not observe statistical difference between the degree of dysplasia and the amount of CD133+ cells (p = 0.899). CD133 immunoexpression showed no association with the OEDs and OSCCs sites. It was observed that nuclear and cytoplasmic immunostaining was more evident with the progression of the malignant process. CONCLUSION: It is suggested that the CD133 cellular localization together with the histopathological criteria of OEDs classification can contribute to provide more concrete indications about the oral carcinogenesis process.

[Value of the expression of Ki-67 and membrane protein glucose transporter GLUT1 in the diagnosis of neoplastic transformation of the oral mucosal epithelium]. / Znachenie ekspressii Ki-67 i membrannogo belka-perenoschika glyukozy GLUT1 v diagnostike neoplasticheskoi transformatsii epiteliya slizistoi obolochki rta.

AIM: to investigate the expression of GLUT1 protein in relation to the proliferative activity of cells and the degree of neoplastic transformation of the epithelium of the oral mucosa (OM). SUBJECTS AND METHODS: OM biopsies from 44 patients were investigated. 14 cases (31.9%) were diagnosed with focal epithelial hyperplasia of the OM; 13 (29.5%) cases had squamous intraepithelial neoplasia (SIN) and 17 (38.6%) had squamous cell carcinoma. Ten intact MO samples were used as a control group. Tissue antigens were determined using mouse monoclonal antibodies to Ki-67 and rabbit antibodies to GLUT1. RESULTS: A direct correlation was found between the intensity of GLUT1 expression on the oral epithelial cell membranes and the proliferative activity of cells with an increase in the phenomena of cellular atypia. CONCLUSION: GLUT1 may be used as a marker for the early diagnosis of neoplastic transformation of stratified squamous epithelial cells.

A GRHL3-regulated repair pathway suppresses immune-mediated epidermal hyperplasia.

Dermal infiltration of T cells is an important step in the onset and progression of immune-mediated skin diseases such as psoriasis; however, it is not known whether epidermal factors play a primary role in the development of these diseases. Here, we determined that the prodifferentiation transcription factor grainyhead-like 3 (GRHL3), which is essential during epidermal development, is dispensable for adult skin homeostasis, but required for barrier repair after adult epidermal injury. Consistent with activation of a GRHL3-regulated repair pathway in psoriasis, we found that GRHL3 is upregulated in lesional skin and binds known epidermal differentiation gene targets. Using an imiquimod-induced model of immune-mediated epidermal hyperplasia, we found that mice lacking GRHL3 have an exacerbated epidermal damage response, greater sensitivity to disease induction, delayed resolution of epidermal lesions, and resistance to anti-IL-22 therapy compared with WT animals. ChIP-Seq and gene expression profiling of murine skin revealed that while GRHL3 regulates differentiation pathways both during development and during repair from immune-mediated damage, it targets distinct sets of genes in the 2 processes. In particular, GRHL3 suppressed a number of alarmin and other proinflammatory genes after immune injury. This study identifies a GRHL3-regulated epidermal barrier repair pathway that suppresses disease initiation and helps resolve existing lesions in immune-mediated epidermal hyperplasia.

Overexpression of Smad proteins, especially Smad7, in oral epithelial dysplasias.

OBJECTIVE: Transforming growth factor ß, via membrane-bound receptors and downstream Smad2-4, 7, can modulate tumorigenesis. Smad2 and Smad3 heterodimerize with Smad4, and the complex migrates to the nucleus to regulate the expression of target genes. Smad7 is a key negative regulator of this signaling pathway. This study aimed to examine Smad2-4, 7 expression and phosphorylated Smad2-3 (p-Smad2-3) in oral epithelial dysplasia and compared it with normal oral mucosa, hyperkeratosis/epithelial hyperplasia and squamous cell carcinoma (SCC). MATERIALS AND METHODS: Immunohistochemical staining of Smad2-4, 7 and p-Smad2-3, was performed for 75 samples of human oral mucosa, including hyperkeratosis/epithelial hyperplasia (n = 20), mild epithelial dysplasia (n = 11), moderate to severe epithelial dysplasia (n = 11), and SCC (n = 43). Normal buccal mucosa samples (n = 9) were also included. RESULTS: A significant increase in Smad7 expression was observed in the ascending order of samples of normal oral mucosa, hyperkeratosis/epithelial hyperplasia/mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, and well-differentiated oral SCC/moderately to poorly differentiated oral SCC. Additionally, significant increases in Smad7 expression were noted as compared with expression of Smad2-4 and p-Smad2-3 in lesions of hyperkeratosis/epithelial hyperplasia, mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, well-differentiated oral SCC, and moderately to poorly differentiated oral SCC. CONCLUSIONS: Our results indicate that Smad proteins, particularly Smad7, in oral epithelial dysplasia and SCC could contribute to the attenuation of Smads anti-proliferative signaling in cancer development. CLINICAL RELEVANCE: Smad7 could be a marker for risk of malignant transformation of oral epithelial dysplasia.

Reappraisal of immunohistochemical profiling of special histological types of breast carcinomas: a study of 121 cases of eight different subtypes.

Histological special types (HST) account for about 25% of breast cancers, and correspond to at least 17 pathological entities. However, their molecular characteristics remain to be determined. The purpose of the present study is to apply the recently used immunohistochemical profiling of HST breast carcinomas (BC), as a surrogate for the molecular subtyping, what could be relevant for therapeutic purposes. One hundred and twenty-one cases were included. Immunohistochemical study was performed on paraffin sections, including markers for oestrogen, progesterone and androgen receptors, keratin 5, HER2/neu, epithelial growth factor receptor, p63 protein, P-cadherin, and Ki-67. Tubular (16 cases), mucinous (27) and papillary (9) types were all categorised as luminal-like A and B. The medullary (21) and metaplastic (10) types corresponded largely to the basal-like tumours (85.7% and 90%, respectively). Cases of the micropapillary type (8) were luminal A (3/8), luminal B (4/8) and HER2 overexpressing (1/8), whereas the apocrine carcinomas (24) presented a heterogeneous profile. The proliferation rate (Ki-67) varied among the types, being the medullary carcinoma subtype with higher proliferation. Comparing the current data with those based on molecular studies, there was good agreement in the classification of the tubular, mucinous and papillary types. Only a partial concordance was achieved for the other types, which may be due to sampling, and to the relatively low frequency of such cases. The present work supports the clinical usage of immunohistochemistry as a surrogate to molecular classification of special types of BC.

[Correlation between the quantity of oral mucosal micronucleus cells and cancerization].

OBJECTIVE: To explore the relationship between micronuclei and malignant development of oral epithelial dysplasia (OED). METHODS: Oral exfoliated cells were smeared from 30 control subjects, 46 patients with oral lichen planus (OLP), 78 patients with oral leukoplakia (OLK) and hyperplasia, 30 patients with OLK and mild OED, 29 patients with OLK and moderate OED, 15 patients with OLK and severe OED and 22 patients with oral squamous cell carcinoma (OSCC). RESULTS: Micronucleus rates declined according to the order of OSCC, OLK and severe OED, OLK and moderate OED, OLK and mild OED, OLK and hyperplasia, OLP and control groups (P<0.001). CONCLUSION: Micronuclei may reflect the progression of oral epithelial malignant changes. It could be used in monitoring cancerization in following up cases regularly.

Serotonin content of normal and inflamed appendix: a possible role of serotonin in acute appendicitis.

The appendix is lined by a mucosa which has many neuroendocrine cells containing serotonin. Local release of serotonin can act as a mediator of inflammation. In this study we explored the serotonin content of the neuroendocrine cells of the appendixes removed for clinical diagnosis of appendicitis. Appendix specimens were divided into three groups: Acute appendicitis (AA), non-appendicitis (NA), and follicular hyperplasia (FH). Normal appendix specimens from patients undergoing elective abdominal surgery were used as the control group (NL). All sections were exposed to proteinase K, incubated with anti-serotonin, chromogranin A, and synaptophysin antibodies, and treated with the LSAB kit. Polygonal cells were seen within the crypt epithelium (enterochromaffin cell, EC) and within the lamina propria (subepithelial neuroendocrine cell, SNC). In AA, only 16 cases (64%) showed serotonin staining in non-destructed glands. There was a significant reduction in the number of ECs in AA compared to the FH (96%), NA (100%) and NL (100%) groups (P<0.001). Chromogranin and synaptophysin immunostaining also showed a significant reduction in the number of ECs in AA compared with the other three groups (P<0.001). SNC serotonin reactivity was lower in the AA group compared with the other groups (p<0.001). The inflamed appendix is markedly depleted of serotonin in the epithelium and lamina propria. Local serotonin release from ECs and SNCs in the appendix may act as an inflammatory mediator in appendicitis and is likely to be the source of raised blood serotonin in AA.

Enhanced levels of glutathione and protein glutathiolation in rat tongue epithelium during 4-NQO-induced carcinogenesis.

High glutathione (GSH) levels are commonly found in oral tumors and are thought to play an important role in tumorigenesis. While posttranslational binding of GSH to cellular proteins (protein glutathiolation) has recently been recognized as an important redox-sensitive regulatory mechanism, no data currently exist on this process during carcinogenesis. Our goal was to determine the effects of 4-nitroquinoline-N-oxide (4-NQO)-induced carcinogenesis on tongue levels of protein-bound and free GSH and related thiols in the rat. Male F-344 rats (6 weeks of age) were administered either 4-NQO (20 ppm) in drinking water or tap water alone (controls) for 8 weeks. Twenty-four weeks after cessation of 4-NQO, squamous cell carcinomas of the tongue were observed in all rats. The levels of both free and bound GSH in tumors, as well as in adjacent tissues, were 2- to 3-fold greater than in tongue epithelium from control rats (p < 0.05). Prior to tumor formation, at 8 weeks after cessation of 4-NQO, hyperplasia, dysplasia and carcinoma in situ were observed in 100%, 25% and 12.5% of 4-NQO-treated rats, respectively. At this early stage of carcinogenesis, levels of free and bound GSH were increased 50% compared with tongue tissues from control rats (p<0.05). Glutathione disulfide (GSSG) levels were also 2-fold greater in tongue tissues from 4-NQO treated vs. control rats (p<0.05). Altogether, these results suggest that protein glutathiolation, together with GSH and GSSG levels, are induced during oral carcinogenesis in the rat possibly as a result of enhanced levels of oxidative stress.

Double immunohistochemical labeling technique applied to different types of cytokeratins in epithelial proliferations of the breast.

The double labeling technique using peroxidase and alkaline phosphatase for immunohistochemistry is well known, but must be adapted according to the antibodies used, fixation, and technical conditions. The technique allows identification on one slide of two antigens that are localized in the same or different cells of the same lesion. The aim of this paper is to describe the adaptation of this technique to cytokeratins of normal mammary tissue and proliferative lesions of the breast.