Cytoplasmic retention of p53 in colorectal cancer cells

30th Congress of the Federation-of-European-Biochemical-Societies (FEBS)/9th IUBMB Conference, Jul 02-07, 2005, Budapest, Hungar

Cytoplasmic retention of p53 in colorectal cancer cells

30th Congress of the Federation-of-European-Biochemical-Societies (FEBS)/9th IUBMB Conference, Jul 02-07, 2005, Budapest, Hungar

Mutant P53 Protein Expression and Antioxidant Status Deficiency in Breast Cancer

It is well recognized that cancers develop and grow as a result of disordered function of tumor suppressor genes and oncogenes, which may be exploited for screening purposes. Extensive evidence indicated tumor suppressor protein p53 as candidate marker for mutation identification. We have investigated mutant p53 protein expression in human breast tumors in relation to antioxidant status deficiency. The study included 100 breast cancer patients. p53 protein expression was evaluated by Western blot assay and immunostaining using a CM-1, DO-7 and Pab240 antibodies. Antioxidant parameters and lipid peroxidation were estimated by biochemical analyses. Western blotting with epitope-specific monoclonal antibody Pab240 strongly suggests that nuclear extracts from breast cancer cells express mutant forms of p53. It is of interest that the mutant forms of p53 overexpression in conjunction with the appearance of nuclear bodies are observed in highly aggressive carcinomas. Expression of isoform Delta p53 (45 kDa) and isoform of similar to 29 kDa were more common in cases with LN metastasis. These studies point out the molecular consequences of oxidative stress (lipid peroxides, LP, p LT 0.001) and antioxidant status deficiency (copper, zinc superoxid dismutase, SOD, p LT 0.001; catalase, CAT, p LT 0.01; glutathione reductase, GR, p LT 0.001; glutathione, GSH, p LT 0.05) and indicate the importance of p53 mutation as the commonest genetic alteration detected in breast cancer cells. The expression of mutant p53 is correlated to increased lipid peroxides (0.346, p LT 0.05) and lowered antioxidant activity of CAT (- 0.437, p LT 0.01) in the breast cancer patients

Mutant P53 Protein Expression and Antioxidant Status Deficiency in Breast Cancer

It is well recognized that cancers develop and grow as a result of disordered function of tumor suppressor genes and oncogenes, which may be exploited for screening purposes. Extensive evidence indicated tumor suppressor protein p53 as candidate marker for mutation identification. We have investigated mutant p53 protein expression in human breast tumors in relation to antioxidant status deficiency. The study included 100 breast cancer patients. p53 protein expression was evaluated by Western blot assay and immunostaining using a CM-1, DO-7 and Pab240 antibodies. Antioxidant parameters and lipid peroxidation were estimated by biochemical analyses. Western blotting with epitope-specific monoclonal antibody Pab240 strongly suggests that nuclear extracts from breast cancer cells express mutant forms of p53. It is of interest that the mutant forms of p53 overexpression in conjunction with the appearance of nuclear bodies are observed in highly aggressive carcinomas. Expression of isoform Delta p53 (45 kDa) and isoform of similar to 29 kDa were more common in cases with LN metastasis. These studies point out the molecular consequences of oxidative stress (lipid peroxides, LP, p LT 0.001) and antioxidant status deficiency (copper, zinc superoxid dismutase, SOD, p LT 0.001; catalase, CAT, p LT 0.01; glutathione reductase, GR, p LT 0.001; glutathione, GSH, p LT 0.05) and indicate the importance of p53 mutation as the commonest genetic alteration detected in breast cancer cells. The expression of mutant p53 is correlated to increased lipid peroxides (0.346, p LT 0.05) and lowered antioxidant activity of CAT (- 0.437, p LT 0.01) in the breast cancer patients

Molecular characterization of hsp90 isoforms in colorectal cancer cells and its association with tumour progression

A key role of hsp90 in the activity of various oncogenic proteins and pathways is currently of intense interest. To clarify the molecular basis of biological behaviour of colorectal cancers we analysed the expression characteristics of hsp90 in cytosolic, nuclear and plasma membranous fractions of cancer cells. As determined by Western blot assay all hsp90 isoforms studied, a (84 kDa), 8 (86 kDa) and hsp90N (75 kDa), were up-regulated and differentially expressed in various stages of colorectal carcinoma. The inducible hsp90 alpha isoform is a component of invasive phenotype of cancer cells thus pointing to the importance of hsp90 alpha for metastasis generation. The expression of hsp90 beta is definitely higher in poorly-differentiated carcinomas than in well-differentiated cancers, suggesting an involvement of hsp90 beta in the inhibition of cancer cell differentiation. Especially, the expression of cytosolic hsp90N isoform in malignant cells points to the possibility that induction or overexpression of hsp90N might be causally related to tumour formation. Hsp90N is the plasma-membrane-associated protein in poorly-differentiated colorectal cancers with metastasis. This suggests that the expression of hsp90N is elevated with progressive dedifferentiation often associated with advanced cancer stages. Hsp90 was exclusively localized in the invasive front in a majority of metastatic cancers as visualized by immunohistochemical study. Consistent with these facts, the frequent expression of hsp90 alpha and hsp90N on the surface of colorectal cancer cells may enable hsp90 to act as a mediator of metastasis generation. The above results indicate more complex roles for hsp90 in colorectal tumourigenesis. In this way, the hsp90 would be at the crossroads of both signalling and cell migration events

Hiperecogenicidade dos vasos talâmicos no recém-nascido prematuro Hyperechogenicity of thalamic vessels in preterm newborn infants

Objetivo: o presente estudo procura avaliar as possíveis patologias que se manifestam associadas à hiperecogenicidade dos vasos talâmicos na ultra-sonografia cerebral, e observar a freqüência com que ocorrem. Métodos: a amostra foi constituída de 206 recém-nascidos prematuros, nascidos no Hospital de Clínicas de Porto Alegre, no período de julho de 1998 a maio de 1999. Todos realizaram a ultra-sonografia cerebral na primeira semana de vida. Foram incluídos no estudo aqueles prematuros que necessitaram de internação hospitalar, e que tiveram o termo de consentimento informado assinado por um dos responsáveis. Foram excluídos aqueles cuja ultra-sonografia cerebral evidenciava sangramento cerebral e/ou malformações congênitas associadas, e os que evoluíram para óbito antes da realização do exame. Resultados: a ultra-sonografia cerebral levou à identificação de 65 recém-nascidos prematuros com hiperecogenicidade dos vasos talâmicos e de 141 recém-nascidos prematuros sem. Conclusão: a forma de apresentação do tipo pélvica ao nascimento, a maior idade gestacional, o maior peso do recém-nascido ao nascimento e a classificação grande para a idade gestacional foram fatores de risco para a ocorrência de hiperecogenicidade dos vasos talâmicos, enquanto a presença de hipertensão materna durante o período de gestação tendeu a ser fator de proteção. Os recém-nascidos que apresentaram crises convulsivas durante o período de internação hospitalar tiveram risco 3,2 vezes maior de ter hiperecogenicidade dos vasos talâmicos, quando comparados aos que não apresentaram crises convulsivas.<br>Objective: the aim of this study is to evaluate possible pathologies associated with hyperechogenicity of thalamic vessels (HETV), which are found on brain ultrasounds (BUS), as well as to observe the frequency of their occurrence. Methods: the sample was composed of 206 preterm newborn infants at Hospital de Clíncas de Porto Alegre (HCPA) from July 1998 to May 1999. All of them were submitted to BUS in the first week of life. Preterm children who needed hospital admission and had a term of informed consent signed up by their guardians were included in this study. Preterm newborn children with BUS showing intracranial hemorrhage and/or associated congenital malformation were excluded from this study. Results: through BUS it was possible to identify 65 preterm newborn children with HETV and 141 preterm newborn children without HETV. Conclusions: we identified the following risk factors for HETV: pelvic presentation, longer gestational period, increased birthweight and big for gestational age classification. On the other hand, mother's hypertension during the gestational period tended to protect infants from HETV. The newborn infants that presented convulsive crises during hospitalization had a 3.2-fold higher risk of having HETV when compared to the ones who did not go through any convulsive crises

Molecular characterization of hsp90 isoforms in colorectal cancer cells and its association with tumour progression

A key role of hsp90 in the activity of various oncogenic proteins and pathways is currently of intense interest. To clarify the molecular basis of biological behaviour of colorectal cancers we analysed the expression characteristics of hsp90 in cytosolic, nuclear and plasma membranous fractions of cancer cells. As determined by Western blot assay all hsp90 isoforms studied, a (84 kDa), 8 (86 kDa) and hsp90N (75 kDa), were up-regulated and differentially expressed in various stages of colorectal carcinoma. The inducible hsp90 alpha isoform is a component of invasive phenotype of cancer cells thus pointing to the importance of hsp90 alpha for metastasis generation. The expression of hsp90 beta is definitely higher in poorly-differentiated carcinomas than in well-differentiated cancers, suggesting an involvement of hsp90 beta in the inhibition of cancer cell differentiation. Especially, the expression of cytosolic hsp90N isoform in malignant cells points to the possibility that induction or overexpression of hsp90N might be causally related to tumour formation. Hsp90N is the plasma-membrane-associated protein in poorly-differentiated colorectal cancers with metastasis. This suggests that the expression of hsp90N is elevated with progressive dedifferentiation often associated with advanced cancer stages. Hsp90 was exclusively localized in the invasive front in a majority of metastatic cancers as visualized by immunohistochemical study. Consistent with these facts, the frequent expression of hsp90 alpha and hsp90N on the surface of colorectal cancer cells may enable hsp90 to act as a mediator of metastasis generation. The above results indicate more complex roles for hsp90 in colorectal tumourigenesis. In this way, the hsp90 would be at the crossroads of both signalling and cell migration events

Expression of heat shock protein 70 (HSP70) in patients with colorectal adenocarcinoma - immunohistochemistry and Western blot analysis

The role of heat shock protein 70 (HSP70) expression has been investigated in various types of tumors. There are only little and controversial data about its clinical relevance in colorectal carcinoma, one of the most common carcinomas observed in humans. In this study we investigated expression of HSP70 in human colonic carcinoma and possible correlation with clinicopathology. To assess patterns (cytosolic and membrane) of HSP70 expression, the 48 surgically removed colorectal adenocarcinomas and 12 normal colonic and rectal mucosal samples were examined by immunohistochemistry and Western-blot. According to results of immunohistochemistry, expression of cytoplasmic HSP72 was significantly higher in colorectal carcinoma compared with normal and adjacent mucosa (p LT 0.01). In addition, there was significant increase in HSP72 expression in lymph node-positive compared to node-nevative group (p LT 0.001). Dukes C2 stage of colonic cancer showed significantly higher immunohistochemical score than Dukes B2 and B1 stage groups (p LT 0.05 i.e. p LT 0.02). There was no relation between expression of HSP72 and degree of tumor differentiation. Using Western blot analyses, we noticed elevated levels of cytosolic HSP70 in colorectal cancer cells compared to normal. Densitometric analysis of blots of plasma membrane HSP70 expression has shown decrease in colorectal cancer cells compared to normal mucosa. According to our results, overexpression of HSP72 in malignant tissues of patients with colorectal carcinoma is related to tumor progression, suggesting that these proteins could play an important role not only in tumorigenesis but also in the development of drug resistance. Further research is necessary to clarify the mechanisms responsible for differential HSP70 expression as well as its definitive role in colorectal cancer

Expression of heat shock protein 70 (HSP70) in patients with colorectal adenocarcinoma - immunohistochemistry and Western blot analysis

The role of heat shock protein 70 (HSP70) expression has been investigated in various types of tumors. There are only little and controversial data about its clinical relevance in colorectal carcinoma, one of the most common carcinomas observed in humans. In this study we investigated expression of HSP70 in human colonic carcinoma and possible correlation with clinicopathology. To assess patterns (cytosolic and membrane) of HSP70 expression, the 48 surgically removed colorectal adenocarcinomas and 12 normal colonic and rectal mucosal samples were examined by immunohistochemistry and Western-blot. According to results of immunohistochemistry, expression of cytoplasmic HSP72 was significantly higher in colorectal carcinoma compared with normal and adjacent mucosa (p LT 0.01). In addition, there was significant increase in HSP72 expression in lymph node-positive compared to node-nevative group (p LT 0.001). Dukes C2 stage of colonic cancer showed significantly higher immunohistochemical score than Dukes B2 and B1 stage groups (p LT 0.05 i.e. p LT 0.02). There was no relation between expression of HSP72 and degree of tumor differentiation. Using Western blot analyses, we noticed elevated levels of cytosolic HSP70 in colorectal cancer cells compared to normal. Densitometric analysis of blots of plasma membrane HSP70 expression has shown decrease in colorectal cancer cells compared to normal mucosa. According to our results, overexpression of HSP72 in malignant tissues of patients with colorectal carcinoma is related to tumor progression, suggesting that these proteins could play an important role not only in tumorigenesis but also in the development of drug resistance. Further research is necessary to clarify the mechanisms responsible for differential HSP70 expression as well as its definitive role in colorectal cancer

Quality of waste water from the dairy industry from the municipality of Kraljevo

Waste waters are becoming an increasing problem of modern society. In the battle for a bigger profit, a large number of industrial enterprises do not pay attention to the consequences that their wastewater to generate environmental quality. On the list of pollutants wastewater significant is the dairy industry. For this reason, the subject of our research was selected wastewater from the dairy industry. The main objective of this paper is to analyze the quality of waste water originating from dairy drive from the municipality of Kraljevo. We analyzed 15 samples of waste water taken from four locations of the manufacturing sites of the dairy industry. This research has shown that some dairy plants, despite legislation, discharging wastewater into the recipient or the public sewerage system, without adequate treatment