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Evaluation of Correlation between mRNA and Protein Expression of Tripeptidyl-Peptidase II: Possible Future Use as a Biomarker for Cancer?Andersson, Daniel January 2013 (has links)
Cancer remains one of the most common causes for death in the world today. Researchers are continuously trying to improve old, and develop new, methods in order to strife this global problem. Much research is being made trying to find new specific biomarkers that can be used to detect and diagnose cancer in an early stage. One candidate protein for possible future use as a biomarker is tripeptidyl-peptidase II (TPPII) which has previously been shown to be up-regulated in Burkitt´s lymphoma. This paper focuses on the expression of TPPII on an mRNA-level to see if there is any difference between expression in human leucocytes from patients with a leukemia diagnosis and a healthy volunteer, in order to evaluate if the expression of TPPII have any future use as a biomarker. Patient samples were analyzed using real time qPCR, to study the expression of mRNA, and Western blot, in order to correlate the mRNA findings with protein expression. Three different cell lines with different characteristics regarding expression and function of TPPII were also used to validate the methods used and for comparison with the patient samples analyzed. A difference in expression of mRNA were seen between the different patient samples, both individually and between larger groups of samples with the same diagnosis, indicating a large individual variation, thus making future use in a clinical setting difficult. However, seeing as only a few samples were analyzed in this study, more research must be done in order to draw any final conclusions.
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ACTH Increases Expression of c-fos, c-jun and β-actin Genes in the Dexamethasone-treated Rat AdrenalsMATSUI, NOBUO, TAKAGI, HIROSHI, FUNAHASHI, HIROOMI, SATOH, YASUYUKI, MIYAMOTO, NORIHIRO, MURATA, YOSHIHARU, IMAI, TSUNEO, SEO, HISAO, OHNO, MOTOTSUGU 08 1900 (has links)
名古屋大学博士学位論文 学位の種類 : 医学博士(論文) 学位授与年月日:平成4年9月22日 大野元嗣氏の博士論文として提出された
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Identificação de potenciais biomarcadores No colesteatoma adquiridoARAÚJO, Juliana Gusmão De 26 December 2012 (has links)
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Previous issue date: 2012-12-26 / O colesteatoma adquirido, mesmo com os conhecimentos acumulados desde sua
primeira descrição, ainda se mantém como um problema de saúde pública distante de ser
solucionado. O entendimento mais profundo da patogênese do colesteatoma é de extrema
importância visto que a natureza desta lesão é destrutiva e causadora de complicações
potencialmente graves. Apesar das teorias propostas e das várias proteínas terem sido
identificados no colesteatoma, a verdadeira etiopatogenia da doença ainda carece de
investigações. Objetivo: Identificar biomarcadores do colesteatoma adquirido utilizando a
plataforma proteômica. Casuística e Métodos: Foram coletadas amostras de colesteatoma e
também fragmento de pele da região retroauricular de 12 indivíduos submetidos a cirurgia
para remoção do colesteatoma. As amostras foram armazenadas em solução salina e
mantidas a -20ºC até pesagem tecidual e extração das proteínas. Eletroforese bidimensional
foi realizada, os géis foram corados com nitrato de prata e suas imagens digitalizadas. Todas
as análises foram realizadas em triplicatas. Os peptídeos extraídos após a digestão do spots
foram levados à espectrometria de massa e os espectros obtidos foram analisados usando o
algoritmo Mascot utilizando os bancos de dados de proteína do NCBI e SwissProt.
Resultados: Dos 393 spots identificados na análise do extrato proteico de colesteatoma
adquirido, apenas 10 estavam dentro dos parâmetros estatísticos aceitáveis pelo algoritmo
Mascot. As principais proteínas detectadas no colesteatoma adquirido foram a cadeia beta do
fibrinogênio, proteína da matriz extracelular 2, actina citoplasmática 1, heparan sulfato
glucosamina 3-O-sulfotransferase 3A1, fator de necrose tumoral alfa induzido proteína 8-like
1, Stanniocalcina-2, lisofosfolipase eosinofílica e OFUT1.Conclusão: Foram identificadas
proteínas envolvidas com a migração celular, regulação da apoptose, vias de sinalização,
hiperproliferação celular, cicatrização e processos inflamatórios. Pudemos, desta maneira,
traçar um perfil proteômico do colesteatoma adquirido. / The acquired cholesteatoma, even with all the knowledge accumulated since its
first description, still remains a public health problem, far from being solved. A deeper
understanding of its pathogenesis is extremely important since it is a destructive lesion that
might cause potentially serious complications. Several proteins have been identified in
cholesteatoma and a few theories were described, however the true etiology of the disease
still needs investigation. Objective: Identify acquired cholesteatoma biomarkers using
proteomics platform. Patients and methods: Cholesteatoma samples were collected and also
a skin fragment of the surgical incision of twelve patients undergoing surgery for
cholesteatoma removal. The samples were stored in saline solution and kept at -20 ° C until
weighing tissue and proteins extraction. Two-dimensional electrophoresis was conducted,
the gels were stained with silver nitrate and their images were digitized. All analyzes were
performed in triplicate. The peptides extracted after spots digestion were taken to mass
spectrometry and the spectra obtained were analyzed using the Mascot algorithm comparing
databases of the NCBI and SwissProt protein. Results: Of the 393 spots identified in the
analysis of protein extracts of acquired cholesteatoma, only 10 were within acceptable
statistical parameters by Mascot algorithm. The proteins detected in acquired cholesteatoma
were fibrinogen beta chain, extracellular matrix protein 2, actin cytoplasmic 1, heparan
sulfate glucosamine 3-O-sulfotransferase 3A1, tumor necrosis factor alpha 8 induced proteinlike
1, stanniocalcin-2, eosinophil lysophospholipase and OFUT1. Conclusion: Proteins
involved in cell migration, regulation of apoptosis, signaling pathways, cellular proliferation,
wound healing and inflammatory processes were identified. We were able to draw a
proteomic profile of acquired cholesteatoma.
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