Expressão da E-caderina em carcinoma de células escamosas e no tumor de células basais de cães

As caderinas compreendem uma classe de moléculas de adesão celular expressa na superfície de todas as camadas epidérmicas. A E-caderina é a principal caderina envolvida na adesão celular epitelial. A redução de sua expressão está envolvida na progressão de alguns tipos de câncer, no potencial metastático e ainda na definição do prognóstico, principalmente nos carcinomas. O carcinoma de células escamosas e o tumor de células basais são neoplasias cutâneas malignas que afetam os cães. O objetivo deste estudo foi avaliar a expressão da E-caderina no carcinoma de células escamosas (n=20) e no tumor de células basais (n=15), buscando-se relacionar sua expressão ao comportamento biológico desses tumores. Os carcinomas de células escamosas apresentaram significativa redução da expressão da molécula comparado aos tumores de células basais, quando avaliado pelo teste de Fisher (P=0,0039). Também foi observado que células neoplásicas mais diferenciadas apresentaram coloração mais intensa que as menos diferenciadas. Em conclusão, sugere-se que a expressão reduzida da E-caderina em tumores cutâneos pode indicar maior poder infiltrativo e consequentemente mau prognóstico na espécie canina.

Avaliação das características histopatológicas e imuno-histoquímicas do fronte de invasão tumoral do carcinoma espinocelular cutâneo de cabeça e pescoço e sua importância no comportamento biológico

Pós-graduação em Patologia - FMB

3D osteoarthritic changes in TMJ condylar morphology correlates with specific systemic and local biomarkers of disease

Objective: To assess 3D morphological variations and local and systemic biomarker profiles in subjects with a diagnosis of temporomandibular joint osteoarthritis (TMJ OA).Design: Twenty-eight patients with long-term TMJ OA (39.9 +/- 16 years), 12 patients at initial diagnosis of OA (47.4 +/- 16.1 years), and 12 healthy controls (41.8 +/- 12.2 years) were recruited. All patients were female and had cone beam CT scans taken. TMJ arthrocentesis and venipuncture were performed on 12 OA and 12 age-matched healthy controls. Serum and synovial fluid levels of 50 biomarkers of arthritic inflammation were quantified by protein microarrays. Shape Analysis MANCOVA tested statistical correlations between biomarker levels and variations in condylar morphology.Results: Compared with healthy controls, the OA average condyle was significantly smaller in all dimensions except its anterior surface, with areas indicative of bone resorption along the articular surface, particularly in the lateral pole. Synovial fluid levels of ANG, GDF15, TIMP-1, CXCL16, MMP-3 and MMP-7 were significantly correlated with bone apposition of the condylar anterior surface. Serum levels of ENA-78, MMP-3, PAI-1, VE-Cadherin, VEGF, GM-CSF, TGF beta b1, IFN gamma g, TNF alpha a, IL-1 alpha a, and IL-6 were significantly correlated with flattening of the lateral pole. Expression levels of ANG were significantly correlated with the articular morphology in healthy controls.Conclusions: Bone resorption at the articular surface, particularly at the lateral pole was statistically significant at initial diagnosis of TMJ OA. Synovial fluid levels of ANG, GDF15, TIMP-1, CXCL16, MMP-3 and MMP-7 were correlated with bone apposition. Serum levels of ENA-78, MMP-3, PAI-1, VE-Cadherin, VEGF, GM-CSF, TGF beta 1, IFN gamma, TNF alpha, IL-1 alpha, and IL-6 were correlated with bone resorption. Published by Elsevier Ltd on behalf of Osteoarthritis Research Society International.

Análise da expressão protéica de moléculas de adesão celular em melanomas cutâneos

Cutaneous melanoma is a malignancy caused by the uncontrolled proliferation of melanocytes. Has been increasing in incidence and mortality over the years. This type of tumor may arise in the skin or any other place where there was migration of neural crest cells. Melanocytes are located in the basal layer of the epidermis forming the epidermal melanin unit, involving a melanocyte to 36 keratinocytes. The proliferation of melanocytes depends on this relationship with the keratinocytes, gap junctions, E-cadherin and desmoglein. The basal membrane is responsible for promoting the integrity of the skin, their adhesion structures shown to be altered in several skin diseases. Some molecules that play the role of adherence in this membrane are the laminins, type IV collagen and type XVII and perlecan. Analysis of protein expression of cell adhesion molecules in melanoma by immunohistochemistry on tissue microarray (TMA). We selected paraffin blocks of patients who were diagnosed with cutaneous melanoma in the period 1995 to 2010, reaching a sample of 124 cases. All these have gone through review of preparation for subsequent histological analysis of tissue microarray (TMA). The protein expression will be analyzed by immunohistochemistry, using markers anti-laminin, anti-collagen type IV and type XVII and anti-perlecan. There was a predominance of female patients, Caucasians and southeastern Brazil. There were frequent in type of extensive superficial ulcers with Breslow depth greater than 4 mm, Clark level IV and mitotic index greater than 6 mitoses per cga. The patients, most of them were female with mean age of 57.13 years and Caucasian. The most frequent histological type was extensive superficial followed by acral lentiginous. Predominated cases very deep, with Breslow thickness greater than 4 mm, Clark IV, ulcerated and on average by 9 cga mitosis, leading to worse prognosis in this sample. It...(Complete abstract click electronic access below)

Neoplasias mamárias de cadelas: expressão gênica e proteica da via Wnt/b-catenina, sua associação com a transição epitélio-mesênquima e prognóstico

Pós-graduação em Medicina Veterinária - FCAV

Chemopreventive activity of systemically administered curcumin on oral cancer in the 4-nitroquinoline 1-oxide model

Curcumin has therapeutic potential in preventing several types of cancer, including colon, liver, prostate, and breast. The goal of this study was to evaluate the chemopreventive activity of systemically administered curcumin on oral carcinogenesis induced by 4-nitroquinolone-1-oxide (4-NQO). A total of 50 male albino rats, Rattus norvegicus, (Holtzman), were divided into five groups (n=10 per group). Four of these groups were exposed to 50 ppm 4-NQO in their drinking water ad libitum for 8 or 12 weeks, two groups were treated with curcumin by oral gavage at 30 or 100 mg/kg per day, and one group was treated with corn oil (vehicle) only. The negative control group was euthanized at baseline. Tongues of all animals were removed after euthanasia and used in the subsequent analysis because the tongue is the primary site of carcinogenesis in this model. Descriptive histological analysis and immunohistochemistry for PCNA, Bcl-2, SOCS1 e-3, and STAT3 were performed to assess the oncogenic process. The gene expression of Vimentin, E-cadherin, N-cadherin, or TWIST1 was assessed using RT-qPCR as a representative of epithelial-mesenchymal transition (EMT) events. The administration of curcumin at 100 mg/kg during the 12 weeks markedly decreased the expression of PCNA, Bcl-2, SOCS1 e -3, and STAT3. Curcumin also minimized the cellular atypia under microscopic analysis and diminished the expression of the genes associated with EMT. These findings demonstrate that the systemic administration of curcumin has chemopreventive activity during oral carcinogenesis induced by 4-NQO.

Evidence of epithelial-mesenchymal transition in canine prostate cancer metastasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chemopreventive Activity of Systemically Administered Curcumin on Oral Cancer in the 4-Nitroquinoline 1-Oxide Model

Curcumin has therapeutic potential in preventing several types of cancer, including colon, liver, prostate, and breast. The goal of this study was to evaluate the chemopreventive activity of systemically administered curcumin on oral carcinogenesis induced by 4-nitroquinolone-1-oxide (4-NQO). A total of 50 male albino rats, Rattus norvegicus, (Holtzman), were divided into five groups (n = 10 per group). Four of these groups were exposed to 50 ppm 4-NQO in their drinking water ad libitum for 8 or 12 weeks, two groups were treated with curcumin by oral gavage at 30 or 100 mg/kg per day, and one group was treated with corn oil (vehicle) only. The negative control group was euthanized at baseline. Tongues of all animals were removed after euthanasia and used in the subsequent analysis because the tongue is the primary site of carcinogenesis in this model. Descriptive histological analysis and immunohistochemistry for PCNA, Bcl-2, SOCS1 e-3, and STAT3 were performed to assess the oncogenic process. The gene expression of Vimentin, E-cadherin, N-cadherin, or TWIST1 was assessed using RT-qPCR as a representative of epithelial-mesenchymal transition (EMT) events. The administration of curcumin at 100 mg/kg during the 12 weeks markedly decreased the expression of PCNA, Bcl-2, SOCS1 e-3, and STAT3. Curcumin also minimized the cellular atypia under microscopic analysis and diminished the expression of the genes associated with EMT. These findings demonstrate that the systemic administration of curcumin has chemopreventive activity during oral carcinogenesis induced by 4-NQO. J. Cell. Biochem. 116: 787-796, 2015. (C) 2014 Wiley Periodicals, Inc.

Androgen therapy reverses injuries caused by ethanol consumption in the prostate: testosterone as a possible target to ethanol-related disorders

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Immunocytochemistry associated with oral exfoliative cytology methodological analysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Epithelial-mesenchymal transition occurs in preneoplastic and neoplastic lesions of canine prostate

Background: The epithelial-mesenchymal transition (EMT) is an essential process in the tumor progression and metastasis. In human prostate carcinoma (PCa), the upregulation of cytokeratin and E-cadherin and down-regulation of vimentin have been associated with aggressive phenotype and poor prognosis. Due to the importance of canine cancer model it was evaluated the immunoexpression of AE1/AE3, E-cadherin and vimentin in canine prostatic lesions. Patients and Methods: A total of 75 prostatic tissues formalin-fixed paraffin embedded from dogs was selected: 10 normal prostatic tissues, 20 benign prostatic hyperplasia (BPH), 25 proliferative inflammatory atrophy (PIA) and 20 PCa. AE1/AE3 was detected with a monoclonal antibody (Invitrogen, 180132) at a 1:300 dilution, applied for 45 min at room temperature (RT). The antibody against Vimentin (V9, Invitrogen) and E-cadherin (NCH-38, Dako cytomatiomn) were monoclonal mouse antibodies, used at a 1:300 and 1:200, respectively, for 45 min at RT. The immunolabelling was performed by a polymer method (Histofine, Nichirei Biosciences,). A negative control was performed for all antibodies by omitting the primary antibody and substituting with Tris-buffered saline. The percentage of C-MYC, E-cadherin, and p63- positive cells per lesion was evaluated according to Prowatke et al. (2007). The samples were scored separately according to staining intensity and graded semi-quantitatively as negative, weakly positive, moderately positive, and strongly positive. The score was done in one 400 magnification field, considering only the lesion, since this was done in a TMA core of 1 mm. For statistical analyses, the immunostaining classifications were reduced to two categories: negative and positive. The negative category included negative and weakly positive staining. Chi-square or Fisher exact test was used to determine the association between the categorical variables. Results: All prostatic normal and BPH tissue were positive for cytokeratin, E-cadherin and negative for vimentin. Similarly, all PIA samples were positive for AE1/AE3. From those samples, 48% (12/25) were also positive for vimentin. 55% of PCa (11/25) was positive for vimentin and among these samples 75% (6/11) was also positive for AE1/AE3 and 45% (5/11) was negative for AE1/AE3. PIA and PCa presented a higher number of vimentin positive cells when compared with normal tissue (p=0.032). E-cadherin expression had no statistical difference among diagnosis groups, but we found a higher number of positive cases, with more than 51% of positive immunostaining in BPH and PIA (81.25% and 78.60% of the cases, respectively) than in PCa (55.55%). Conclusion: The carcinogenesis process regarding prostatic epithelial cells in dogs showed higher vimentin protein expression associated with concomitant loss of the cytokeratin and E-cadherin, similar in humans.

Increased cyclooxygenase-2 and vascular endothelial growth factor protein expression is associated with canine prostatic carcionogenesis process

Background: COX-2 is one of the most important prostaglandin involved in urologic cancer and seems to be associated with tumor progression, invasion, and metastasis. In addition, several effects have been reported for VEGF, including inducing angiogenesis, promoting cell migration, and inhibiting apoptosis. COX2 and VEGF up-regulation have been reported in human prostate cancer. Due to the importance of canine natural model for prostate cancer, the aim of this study was to evaluate COX-2 and VEGF protein expression in canine carcinogenic process. Material and Methods: Seventy-four prostatic tissues from dogs were selected to be evaluated for protein expression by immunohistochemistry (IHC), including: 10 normal prostatic tissues, 20 benign prostatic hyperplasias (BPH), 25 proliferative inflammatory atrophies (PIA) and 20 prostatic carcinomas (PCa). COX-2 and VEGF were detected using the monoclonal antibody CX-294 (1:50 dilution, Dako Cytomation and sc-53463 (1:100 dilution, Santa Cruz), respectively. The immunolabelling was performed by a polymer method (Histofine, Nichirei Biosciences). All reaction included negative controls by omitting the primary antibody. The percentage of C-MYC, E-cadherin, and p63- positive cells per lesion was evaluated according to Prowatke et al. (2007). The samples were scored separately according to staining intensity and graded semi-quantitatively as negative, weakly positive (1), moderately positive, and strongly positive. The score was done in one 400 magnification field, considering only the lesion, since this was done in a TMA core of 1 mm. For statistical analyses, the immunostaining classifications were reduced to two categories: negative and positive. The negative category included negative and weakly positive staining. Chi-square or Fisher exact test was used to determine the association between the categorical variables. Results: The COX-2 protein expression was elevated in the cytoplasm of the canine PCa and PIA compared to normal prostate (p=0.002). VEGF protein expression was increased in 94.75% of the PCa and 100% of the PIA compared with to normal prostate (p = 0.001). No difference was found when compared normal prostate with BPH. Conclusions: This study has demonstrated that the carcinogenesis of canine prostatic tissue may be related to gain of COX-2 and VEGF protein expression.

Expressão gênica e proteica de APC, E-caderina, β-catenina e Caveolina-1 no processo carcinogênico da próstata canina e suas metástases

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Inibição da metástase via transição epitélio-mesenquimal por shRNA, metformina e Y27632 em neoplasia mamária

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)