Activity of selenium on cell proliferation, cytotoxicity, and apoptosis and on the expression of CASP9, BCL-XL and APC in intestinal adenocarcinoma cells

Intestinal cancers are correlated with diet. Thus, determining and understanding nutrient-genome interactions is important. The present work assessed the action of the oligoelement selenium on cell proliferation, cytotoxicity, and in situ apoptosis induction and on the expression CASP9, BCL-XL and APC genes in intestinal adenocarcinoma cells (HT29). HT29 cells were cultured and treated with selenium at concentrations of 5, 50 and 500 ng/mL with or without the damage-inducing agent doxorubicin. These cells were then evaluated for cytotoxicity (MTT), cell proliferation and in situ apoptosis induction. To evaluate gene expression, only the cells treated with 500 ng/mL of selenium were used. RNA was extracted from these cells, and the expressions of CASP9, BCL-XL and APC were analyzed by the RT-PCR method. The GAPDH gene was used as a reference gene. The MTT assay showed that selenium was not cytotoxic at any of the concentrations tested. The cell proliferation assay showed that selenium did not interfere with cell proliferation at the three concentrations tested. In contrast, when the three concentrations were combined with doxorubicin, a significant decrease in the proliferation rate was observed. The apoptosis rate was significantly increased in the selenium (500 ng/mL) and doxorubicin group. CASP9 expression was increased and BCL-XL expression decreased in the selenium (500 ng/mL) and doxorubicin group. APC was significantly increased in the selenium group alone. These results show that selenium increases apoptosis, especially when it is associated with a damage-inducing agent. Also, selenium has an important role in the expression of the APC gene, which is related to cell cycle regulation. (C) 2011 Elsevier B.V. All rights reserved.

Atividade do micronutriente selênio na cinética de proliferação celular, citotoxicidade, indução de apoptose e expressão dos genes CASP9, BCL-XL e APC em células HT29

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estrogen-Responsive Genes Overlap with Triiodothyronine-Responsive Genes in a Breast Carcinoma Cell Line

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

C-2, N-dimethylbenzylamine cyclopalladated compounds: evaluation of cytotoxic, mutagenic and antitubercular activities

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

Undifferentiated pulmonary adenocarcinoma of clear cells associated to hypertrophic osteopathy in a dog

Background: Most of the primary pulmonary tumors in dogs are malignant and from epithelial origin, being bronchioalveolar tumors more prevalent. Adenocarcinoma of clear cells, however, is a very rare pulmonary tumor and its origin is still unknown. It is related to several clinical abnormalities, including hypertrophic osteopathy, an unusual paraneoplastic syndrome characterized by a periosteal reaction along the shaft of long bones. Because of the unusual presentation of the pulmonary adenocarcinoma, the aim of this study was to describe the radiographic, histopathological, and immunohistochemical fi ndings of a dog affl icted with hypertrophic osteopathy secondary to an undifferentiated pulmonary adenocarcinoma of clear cells. Case: A 12-year-old, 45 kg, not castrated male Great Dane dog was presented with painful swelling of all four limbs and moderate respiratory distress. Radiographic examination and computed tomography of the limbs showed palisade-like periosteal bone proliferation involving radius, ulna, femur, patella, tibia, fi bula, tarsus, metacarpal, metatarsal and digits, suggesting hypertrophic osteopathy. Radiographic examination and computed tomography of the lungs also showed a round mass well delimited localized in the right diaphragmatic lobe. A lobectomy of the right diaphragmatic lobe and partial lobectomy of accessory lobe were performed. A poorly differentiated clear squamous cell carcinoma was diagnosed by histological examination. An immune-panel of CK5/CK6, CK7, p63 and TTF-1 was used for immunophenotyping. Immunostaining was weakly positive for CK5/CK6 and negative to all others. Therefore, the diagnosis was poorly differentiated clear cell adenocarcinoma. The dog showed improvement in clinical signs seven days after surgery. One month postoperatively, radiographic examination of the limbs showed less intense periosteal reaction and initiation of bone remodeling. Discussion: Primary pulmonary tumors are considered very infrequent in small animals, but its true incidence rate is dif- fi cult to establish in animal populations. The histological origin of the tumor in the present case, as verifi ed in the literature, is not well established by histological analysis. In these situations, the immunohistochemistry panel may be useful. The modifi cation of the diagnosis between histological analysis and by immunohistochemistry, among other factors, might be due to transdifferentiation from one phenotype to another at various stages in the neoplastic process. The clear cell appearance observed in this case may be verifi ed in all types of carcinoma due to intracellular accumulation of glycogen, most of which is dissolved during the preparation of paraffi n sections. This uncommon neoplasm apparently did not infl uence the radiographic or tomographic fi ndings of the hypertrophic osteopathy in the present case. The frequency of metastases depends on the histological type of the tumor, being common in the pulmonary adenocarcinoma and usually to tracheobronchial lymph nodes and pulmonary parenchyma. Although in this case the imaging studies did not show metastases to other pulmonary lobes, the histological exams showed metastatic lesions that may be associated to the dog’s death after the surgery.

Genotyping of AR and PSA polymorphisms in a patient with Klinefelter syndrome, non-Hodgkin lymphoma, and adenocarcinoma of the prostate

It has been hypothesized that the AR (androgen receptor) gene binds the two PSA (prostate-specific antigen) alleles with differing affinities and may differentially influence prostate cancer risk. In this article, we report a case of adenocarcinoma of the prostate in a 56-year-old man with Klinefelter syndrome (47,XXY) and non-Hodgkin lymphoma, as well as the AR and PSA genotype. AR and PSA gene polymorphisms were analyzed by polymerase chain reaction-based methods using DNA from peripheral white blood cells and the prostate cancer. We determined the methylation status of the AR gene on the X chromosome. The patient presents with the AG genotype for the ARE-I (androgen response element) region of the PSA gene. We detect the presence of two short AR alleles with 19 and 11CAG repeats each. Unmethylated alleles were demonstrated for both. The shorter allele was inactive in more than 60% of total DNA in both control blood and prostate cancer cells. The presence of short AR alleles and the G allele of the PSA gene may contribute to the development of prostate cancer in a 47,XXY patient. (C) 2004 Elsevier B.V. All rights reserved.

Polysaccharide fraction of Agaricus brasiliensis avoids tumor-induced IL-10 production and changes the microenvironment of subcutaneous Ehrlich adenocarcinoma

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

Adenocarcinoma of the parotid salivary gland in a cow

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Deletion (1)(q12) and double minutes in a metastatic adenocarcinoma of the prostate

Cytogenetic analysis after short-term cultures were performed on an invasive and moderately-differentiated prostatic adenocarcinoma. The results showed a normal male chromosomal complement in most metaphases examined. Furthermore, several abnormalities were found, including del(1)(q12), double minute and ring chromosomes, acentric fragments, triradial figures, and near-tetraploid cells. (C) Elsevier B.V., 1999. All rights reserved.

Anatomy of smooth muscle cells in nonmalignant and malignant human prostate tissue

Differently graded areas of human prostate adenocarcinoma were examined after Masson's trichrome staining or immunohistochemistry for smooth muscle alpha-actin, type IV collagen and laminin. In addition, the ultrastructure of the prostatic smooth muscle cells (SMC) during glandular proliferation and epithelial invasion in selected tumors was studied. The SMC formed a thick layer below the epithelial structures in unaffected areas and were closely associated with each other in homotypic interactions. As the tumor grade increased, the SMC gradually lost interactions with each other and became atrophic. With the growth of the epithelial compartment, the SMC initially segregated to the tumor periphery and the intercellular spaces increased. In high grade tumors, the epithelial cancer cells invaded the spaces between the SMC. Immunohistochemical analysis of the basal membrane revealed increased disruption of the usually thick basal membrane, which became thinner and faintly stained with each of the antibodies used. We conclude that most SMC become atrophic following epithelial invasion in human tumors and that degradation of the basal membrane is an important factor in this process. At the ultrastructural level, different SMC phenotypes occur in prostatic tissues during epithelial invasion. Interconversion between these phenotypes is suggested and a probable relationship among them is proposed.

Characterization of the cellular component of polymorphous low-grade adenocarcinoma by immunohistochemistry and electron microscopy

In order to characterize the cellular component of the polymorphous low-grade adenocarcinoma (PLGA) of the salivary gland, a morphological and immunohistochemical study was carried out. Thirty cases of PLGA were studied by light microscopy and immunohistochemistry and five cases by transmission electron microscopy (TEM). The expression of cytokeratins (CKs) 7,8,10,13,14,18,19, vimentin and muscle-specific actin (MSA) was investigated through the streptavidin-biotin method. The majority of tumor cells stained for vimentin, CKs 8,18 and 7. CK 14 was positive in most cells of the papillary and trabecular sub-types. Although the expression of CKs 8,18 and 14 varied among the tumors sub-types, a straight relationship between each histologic pattern and the CK expression could not be delineated. MSA was reactive in only three tumors while CKs 10 and 13 were not detected in any tumor studied. The absence of MSA and the expression of CKs 8,18 and 7, in most of the tumor cells, lead to the hypothesis that myoepithelial cells are not the major cellular component of the PLGA. TEM revealed cells exhibiting microvilli and variable amounts of secretory granules, some of them suggesting an excretory activity. The presence of CKs 8, 18 and 7, added to the secretory granules, indicates that PLGA originates from cells located at the acinar-intercalated duct junction. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

Characterizing subpopulations of neoplastic cells in serous effusions - the role of immunocytochemistry

OBJECTIVE: To analyze the role of immunochemistry in serous effusions.STUDY DESIGN: We analyzed cell blocks of 18 pleural and 18 peritoneal effusions diagnosed as malignant (18), benign (14) and suspicious (4). They were immunostained by the avidin-biotin complex method with a panel of four monoclonal antibodies-CEA, Ber-EP4, LeuM1 (CD15) and r53-and for lectins (Ulex europaeus) UEA-1, ConA and ConBr. RESULTS: Seventeen of the 18 cases of adenocarcinoma were positive for CEA (95%), 12 (66.6%)for Bev-EP4, 11 (61 %)for CD15 nild 11 (62 %)for p53. Twelve of the 18 (66.6%) were positive for UEA-1, CEA, Ber-EP4 and CD15. UEA-1 did not react with mesothelial cells. p53 Gave a positive reaction in only one case, reactive mesothelial cells. ConA and ConBr reacted indiscriminately with benign and malignant cells; thus, it was not useful in distinguishing between these cells.CONCLUSION: In this context no antibody used alone is reliable for corroborating a diagnosis, but the selective use of a small panel of three markers (CEA, Ber-EP4 and LeuM1) can be very useful in solving diagnostic difficulties in the cytodiagnosis of serous effusions.

Adenocarcinoma polimorfo de baixo grau de malignidade do tipo papilífero. Estudo morfológico e imuno-histoquímico.

Two cases of polymorphous low-grade adenocarcinoma of the papillary type, from minor salivary glands were studied by light microscopy and immunohistochemistry. One case exhibited a predominance of the papillary pattern, whereas the other presented the following patterns of histological appearance: papillary, solid, pseudocystic and tubular. Utilizing the peroxidase-antiperoxidase (PAP) method, the intermediate filament vimentin, keratin and S100 protein were observed in tumor cells. The immunohistochemical analysis revealed two types of neoplastic cells: myoepithelial and luminal.

Adenocarcinoma de cólon com diferenciação neuro-endócrina e escamosa. Relato de caso.

A tubular adenocarcinoma of the colon with solid area composed by small cells that was found by immunohistochemistry study using antibody to neuron-specific enolase (NSE) to possess neuroendocrine differentiation is related. In another areas of the tumor were visualized keratinizing squamous cells. The presence of neuro-endocrine and squamous cells features provide further evidence that neoplastic colonic cells have the capacity for multi-directional differentiation. The implications of this combination in relation to theories of tumor origin and differentiation and the prognostic significance of neuro-endocrine cells in malignant neoplasms of the gastrointestinal tract are discussed.

Adenocarcinoma in females detected in serous effusions: Cytomorphologic aspects and immunocytochemical reactivity to cytokeratins 7 and 20

OBJECTIVE: To investigate the usefulness of assessing the immunoreactivity of cytokeratins 7 (CK7) and 20 (CK20) as well as several cytomorphologic parameters in effusions with metastatic adenocarcinomas in the search for the primary site of the tumor. STUDY DESIGN: From the files of the Pathology Department, A. C. Camargo Hospital, we studied cytologic smears from 73 metastatic adenocarcinomas originally from the breast, 63 from the ovary, 40 from the lung and 32 from the stomach, looking for morphologic parameters that could have discriminant potential in suggesting the primary site in a routine situation, including intranuclear inclusions, prominent nucleoli, mitosis, signet-ring cells, psammoma bodies, nuclear crease, binucleation and multinucleation, papillary features, acinar profile (including ball cells) and single cells. Immunoreactions were performed with monoclonal antibodies to CK7 (OV-TL 12/30 and CK20 (Ks 20.8) and included morphologic analysis. Both analyses were studied in a blind fashion regarding the primary site of the tumors. RESULTS: Positivity ratios for breast, ovary, stomach and lung cases were 67.6%, 63.5%, 29.7% and 45.5%, respectively, for CK7 and 17.2%, 15.8%, 13.5% and 32.2%, respectively, for CK20. Discriminant analysis of morphologic and immunocytochemical parameters had an error rate of 42.9% in recognizing the primary site and a Wilk's lambda of .7290. CONCLUSION: The more efficient parameter with discriminant function was the papillary appearance showed by CK7, which should be used in further studies with a similar scope. The set of parameters used in this study were insufficient to discriminate the primary site of female adenocarcinomas in effusions with significant accuracy.