Sueli Nonogaki

25(OH)D3 and 1,25(OH)2D3 serum concentration and breast tissue expression of 1α-hydroxylase, 24-hydroxylase and Vitamin D receptor in women with and without breast cancer

1,25(OH)2D3 is an antiproliferative agent that may inhibit proliferation of breast cancer (BC) cells in vitro and BC development in animals. Epidemiological studies have shown a high incidence of BC in people less exposed to solar rays. To unravel the role of Vitamin D3 in BC patients, we have investigated serum levels of 25(OH)D3 and its active form 1,25(OH)2D3 as well as tissue expression of 1alpha-hydroxylase, 24-hydroxylase, and Vitamin D-receptor (VDR), determined by semiquantitative RT-PCR, in 88 Brazilian BC patients and 35 women without cancer (submitted to mammoplasties or resection of benign lesions). Median age of women with and without cancer was 51 and 46 years, respectively, and the majority of BC patients were classified as clinical stage II (67%). Although no differences in 25(OH)D3 serum concentration were found, 1,25(OH)2D3 (40+/-21 pg/ml) levels in BC patients were lower than in women without cancer (53+/-23). Our results indicate that 24-hydroxylase, VDR and 1alpha-hydroxylase mRNA tissue expression is similar in both groups and no correlation between 24-hydroxylase, 1alpha-hydroxylase, and VDR expression in breast tumors was found. A low 1,25(OH)2D3 serum concentration seems to be associated to breast cancer, however, the mechanism involved in this regulation is still unclear.

Platelet-activating factor receptor (PAF-R)-dependent pathways control tumour growth and tumour response to chemotherapy

BackgroundPhagocytosis of apoptotic cells by macrophages induces a suppressor phenotype. Previous data from our group suggested that this occurs via Platelet-activating factor receptor (PAF-R)-mediated pathways. In the present study, we investigated the impact of apoptotic cell inoculation or induction by a chemotherapeutic agent (dacarbazine, DTIC) on tumour growth, microenvironmental parameters and survival, and the effect of treatment with a PAF-R antagonist (WEB2170). These studies were performed in murine tumours: Ehrlich Ascitis Tumour (EAT) and B16F10 melanoma.MethodsTumour growth was assessed by direct counting of EAT cells in the ascitis or by measuring the volume of the solid tumour. Parameters of the tumour microenvironment, such as the frequency of cells expressing cyclo-oxygenase-2 (COX-2), caspase-3 and galectin-3, and microvascular density, were determined by immunohistochemistry. Levels of vascular endothelium growth factor (VEGF) and prostaglandin E2 (PGE2) were determined by ELISA, and levels of nitric oxide (NO) by Griess reaction. PAF-R expression was analysed by immunohistochemistry and flow cytometry.ResultsInoculation of apoptotic cells before EAT implantation stimulated tumour growth. This effect was reversed by in vivo pre-treatment with WEB2170. This treatment also reduced tumour growth and modified the microenvironment by reducing PGE2, VEGF and NO production. In B16F10 melanoma, WEB2170 alone or in association with DTIC significantly reduced tumour volume. Survival of the tumour-bearing mice was not affected by WEB2170 treatment but was significantly improved by the combination of DTIC with WEB2170. Tumour microenvironment elements were among the targets of the combination therapy since the relative frequency of COX-2 and galectin-3 positive cells and the microvascular density within the tumour mass were significantly reduced by treatment with WEB2170 or DTIC alone or in combination. Antibodies to PAF-R stained the cells from inside the tumour, but not the tumour cells grown in vitro. At the tissue level, a few cells (probably macrophages) stained positively with antibodies to PAF-R.ConclusionsWe suggest that PAF-R-dependent pathways are activated during experimental tumour growth, modifying the microenvironment and the phenotype of the tumour macrophages in such a way as to favour tumour growth. Combination therapy with a PAF-R antagonist and a chemotherapeutic drug may represent a new and promising strategy for the treatment of some tumours.

Role of Fos-related antigen 1 in the progression and prognosis of ductal breast carcinoma.

Logullo A F, Stiepcich M M Á, de Toledo Osório C A B, Nonogaki S, Pasini F S, Rocha R M, Soares F A & Brentani M M
(2011) Histopathology 58, 617–625
Role of Fos‐related antigen 1 in the progression and prognosis of ductal breast carcinoma

Prion protein ablation increases cellular aggregation and embolization contributing to mechanisms of metastasis

Cellular Prion Protein (PrPC) is a cell surface protein highly expressed in the nervous system, and to a lesser extent in other tissues. PrPC binds to the extracellular matrix laminin and vitronectin, to mediate cell adhesion and differentiation. Herein, we investigate how PrPC expression modulates the aggressiveness of transformed cells. Mesenchymal embryonic cells (MEC) from wild‐type (Prnp+/+) and PrPC‐null (Prnp0/0) mice were immortalized and transformed by co‐expression of ras and myc. These cells presented similar growth rates and tumor formation in vivo. When injected in the tail vein, Prnp0/0ras/myc cells exhibited increased lung colonization compared with Prnp+/+ras/myc cells. Additionally, Prnp0/0ras/myc cells form more aggregates with blood components than Prnp+/+ras/myc cells, facilitating the arrest of Prnp0/0ras/myc cells in the lung vasculature. Integrin αvβ3 is more expressed and activated in MEC and in transformed Prnp0/0 cells than in the respective Prnp+/+ cells. The blocking of integrin αvβ3 by RGD peptide reduces lung colonization in transformed Prnp0/0 cells to similar levels of those presented by transformed Prnp+/+ cells. Our data indicate that PrPC negatively modulates the expression and activation of integrin αvβ3 resulting in a more aggressive phenotype. These results indicate that PrPC may have main implications in modulating metastasis formation.

Evaluation of prognostic factors in stage IIA breast tumors and their correlation with mortality risk

ABSTRACT: Breast tumors exhibit extensive molecular and clinical heterogeneity. One of the most utilized breast carcinoma classifications is based on its molecular aspects and subdivides breast cancer into five major groups based on the expression of certain genes. In this study, we evaluated which factors are important in determining a prognosis after 5 years of follow-up for patients with clinical stage IIA breast tumors. We took into consideration the different phenotypes (luminal A luminal B HER-2 overexpression, basal and triple-negative), various epithelial-mesenchymal (EMT) molecular markers and adhesion molecules (E-cadherin, P-cadherin, N-cadherin, vimentin, twist snail and slug) and NOS-2, in addition to clinical and demographic data, tumor characteristics and treatment types. METHODS: The study population consisted of 82 patients with breast cancer. We analyzed eight molecular markers by immunohistochemistry on tissue microarrays containing breast tumor specimens from patients with ten years of follow-up, and we classified each tumor according to its estrogen receptor, progesterone receptor and HER-2 expression. We then placed the tumor into one of the above categories. RESULTS: The presence of several clinical and demographic factors, various histopathologies, treatment forms and several immunohistochemical markers were not associated with a worse prognosis for group IIA patients. The factors that were associated with a mortality risk were the triple-negative (odds ratio (OR) = 11.8, 95% confident interval (CI) = 2.0-70.3, P = 0.007) and basal (OR = 18.4, 95% CI = 1.8-184.7, P = 0.013) phenotypic patterns. CONCLUSIONS: The EMT markers and NOS-2 were not mortality risk factors. Basal and triple-negative phenotypic patterns were related to a higher mortality risk in patients with stage IIA tumors.

The expression of metaloproteinases-2 and -9 is different according to the patterns of growth and invasion in squamous cell carcinoma of the penis

Squamous cell carcinoma (SCC) of the penis is characterized by different patterns of growth and local invasion. The matrix metalloproteinases (MMPs) is a family of proteolytic enzymes that are involved in the degradation of extracellular matrix to allow the migration of tumor cells. The present study examined whether the expression of MMP-2 and -9 is correlated with the patterns of tumor growth and invasion in penile SCC. The expression of MMP-2 and -9 was examined immunohistochemically in samples of 115 patients. The cases were divided in three groups according to the patterns of growth and invasion: group 1, exophytic growth and pushing pattern of invasion; group 2, endophytic growth and invasion in large sheets of cells; and group 3, endophytic growth and invasion in small group or isolated cells. Tumors with MMP-2 and -9 overexpression are deeply invasive and present an invasion pattern of small groups of cells. Also, expression of MMP-2 changed from membrane to cytoplasm in invasive tumors, maybe representing activation of MMP-2. These findings allow us to conclude that the less differentiated tumors, which are more invasive and with a pattern of invasion in small group of cells, are associated with the overexpression of MMPs.

P53 overexpression in epidermoid carcinoma of the head and neck.

UNLABELLED The theory of field cancerization in tumors of the head and neck reflects the complex oncogenesis that occurs in this region. The mechanisms that control cell proliferation at the molecular level in epidermoid carcinomas (ECs) of the upper aerodigestive tract are still unclear. Mutations in p53 are the genetic alterations most often detected in ECs of the head and neck and seem to contribute actively to the carcinogenic process triggered by p53 as a tumor-suppressor gene and to its association with tobacco. The objective of the present study was to investigate the expression of p53 protein in epidermoid head and neck carcinomas by immunohistochemistry and its immunohistochemical correlation with other prognostic factors. The study was conducted on 63 consecutive ECs cases not submitted to previous treatment. Specimens of the tumor and of the normal adjacent mucosa were collected during surgery and submitted to immunohistochemical reaction for the determination of the expression of anti-protein p53 antibody (M7001 DAKO A/S, Denmark). Anatomo-clinical and demographic data were not significantly correlated with the presence of lymph node metastases or p53 expression in the tumor or in the adjacent normal mucosa. Tumor localization in the larynx was significantly correlated with p53 expression. Histological grading as grades I, II, III and IV was correlated with significant p53 expression (p = 0.025). CONCLUSIONS 1) in the studied material obtained from 63 cases of head and neck ECs, we detected a 48 percent rate of immunohistochemically detectable p53 overexpression; 2) we did not detect a relationship between demographic patient data and p53 expression in the tumor or in the normal adjacent mucosa; 3) p53 overexpression was significantly more frequent in ECs material from the larynx: and 4) The presence of 12 cases with p53 overexpression in the normal adjacent mucosa and with a p53-negative tumor is in agreement with the theory of field cancerization. Follow-up of this patient series for a longer period of time will permit a better analysis of these values.

Expressão imuno-histoquímica de c-erb-B2 e p53 em carcinomas gástricos

INTRODUCAO: Em nosso meio, os carcinomas gastricos ainda sao neoplasias bastante frequentes e responsaveis por altas taxas de mortalidade. Recentemente, tem-se demonstrado a expressao de p53 e a amplificacao do gene c-erb-B2 nos carcinomas gastricos. A relevância e o significado biologico destas alteracoes ainda nao foram totalmente estabelecidos. OBJETIVO: Estudar as expressoes imuno-histoquimicas de p53 e c-erb-B2 em 482 casos de carcinomas gastricos. MATERIAL E METODOS: Foram construidos tres blocos de tissue microarray (TMA) utilizando-se duplicatas de 482 casos de carcinomas gastricos. Os cortes foram corados por hematoxilina e eosina (HE), tendo sido feita pesquisa para p53 e c-erb-B2. Foram considerados positivos para p53 os casos com marcacao nuclear em mais de 10% das celulas tumorais. Para o c-erb-B2 foram considerados positivos os casos com marcacao de membrana completa em mais de 10% das celulas tumorais. RESULTADOS: A expressao de p53 e c-erb-B2 foi observada em 30% e 12% dos casos, respectivamente. Em relacao aos tipos histologicos observou-se correlacao entre os carcinomas do tipo intestinal e a expressao de c-erb-B2 (p < 0,001). A expressao de p53 foi mais frequente nos carcinomas com mais de 5cm de diâmetro (p = 0,036). Nao foram observadas alteracoes nas curvas de sobrevida dos pacientes em relacao as expressoes desses marcadores. CONCLUSAO: Em nosso meio, carcinomas gastricos do tipo intestinal sao mais frequentemente positivos para c-erb-B2 nos tipos intestinais do que nos difusos. A expressao de p53 esta associada ao tamanho tumoral. A tecnica do TMA e valida e eficiente para o estudo de marcadores imuno-histoquimicos, com forte correlacao com os cortes tradicionais de representacao do tumor.

Análise imuno-histoquímica das sintases do óxido nítrico em adenocarcinomas gástricos

INTRODUCTION: Nitric oxide is an important bioactive and signaling molecule that mediates a diverse array of actions such as vasodilatation, neurotransmission, and iron metabolism. Also, it can act as a carcinogen. Recent studies have examined the expression and activity of the NOS isoforms in several human cancers. OBJECTIVES: To investigate the expression of nitric oxide synthases in gastric carcinomas and correlate the results. MATERIAL AND METHODS: The immunohistochemistry expression of constitutive and inducible nitric oxide synthases (NOS-1, NOS-2 e NOS-3) were evaluated in 128 cases of gastric cancer classified according to Lauren system. RESULTS: The rate of expression of NOS-1 was 92 (70%) of the 12 cases, NOS-2 was 36 (30%) and NOS-3 was 54 (42%) of the cases. The expression of NOS-3 was associated with the intestinal type of carcinoma and deeply invasive tumors showed high rate of expression of NOS-2. CONCLUSION: There is high expression of all the isoforms of nitric oxide synthases in gastric cancer; the constitutive isoforms show higher expression than the inducible forms. The high expression of the inducible form in deeply invasive tumors is related with tumoral progression and dissemination in the gastric mucosa.

Expressão do p53 no carcinoma epidermóide do lábio

BACKGROUND: To assess the p53 expression in squamous cell carcinoma (SCC) of the lip. METHODS: Immunohystochemical study for samples fixed in formaline and paraphin in bloc stained with anti-p53 antibodies through Streptavidina-Biotina-Peroxidase. For statistical analysis the Fisher Test was employed for group differences (p<0.10). RESULTS: The p53 expression were positive in 87.50% of well differentiated SCC, 60.0% of moderately differentiated tumors, 91.67% of poorly differentiated; for surgical margins, 94.23% were negative and 5.77% positive, with an association between differentiation degree and p53 expression (p=0.05). CONCLUSION: The p53 expression was significant in SCC of the lip and negative for surgical margins, but it is not determinant of changing in surgical paradigm.