RESUMO
OBJECTIVE: To explore the differentially expressed proteins in normal cervix, cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) tissues by differential proteomics technique. METHODS: Cervical tissues (including normal cervix, CIN and CSCC) were collected in Department of Gynecologic Oncology of Beijing Obstetrics and Gynecology Hospital. Two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and DeCyder software were used to detect the differentially expressed proteins. Matrix-assisted laser desorption/ionization-time-of-flight tandem mass spectrometry (MALDI-TOF/TOF MS) was used to identify the differentially expressed proteins. Western blot (WB) and immunohistochemistry (IHC) were performed to validate the expressions of selected proteins among normal cervix, CIN and CSCC. RESULTS: 2-D DIGE images with high resolution and good repeatability were obtained. Forty-six differentially expressed proteins (27 up-regulated and 19 down-regulated) were differentially expressed among the normal cervix, CIN and CSCC. 26 proteins were successfully identified by MALDI-TOF/TOF MS. S100A9 (S100 calcium-binding protein A9) was the most significantly up-regulated protein. Eukaryotic elongation factor 1-alpha-1 (eEF1A1) was the most significantly down-regulated protein. Pyruvate kinase isozymes M2 (PKM2) was both up-regulated and down-regulated. The results of WB showed that with the increase in the severity of cervical lesions, the expression of S100A9 protein was significantly increased among the three groups (P = 0.010). The expression of eEF1A1 was reduced but without significant difference (P = 0.861). The expression of PKM2 was significantly reduced (P = 0.000). IHC showed that protein S100A9 was mainly expressed in the cytoplasm, and its positive expression rate was 20.0 % in normal cervix, 70.0 % in CIN and 100.0 % in CSCC, with a significant difference among them (P = 0.006). eEF1A1 was mainly expressed in the cell plasma, and its positive expression rate was 70.0 % in normal cervix, 73.3 % in CIN and 60.0 % in CSCC tissues, without significant difference among them (P = 0.758). PKM2 was mainly expressed in the cell nuclei, and its positive expression rate was 100.0 % in normal cervix, 93.3 % in CIN and 75.0 % in CSCC tissues, showing a difference close to statistical significance (P = 0.059) among them. CONCLUSIONS: There are differentially expressed proteins among normal cervix, CIN and CSCC. S100A9, eEF1A1 and PKM2 may become candidate markers for early diagnosis of cervical cancer and new targets for therapy. It also provides a basis for further studies of the mechanism for CIN developing to CSCC.
Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/metabolismo , Colo do Útero/metabolismo , Displasia do Colo do Útero/metabolismo , Neoplasias do Colo do Útero/metabolismo , Adulto , Western Blotting , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/cirurgia , Estudos de Casos e Controles , Colo do Útero/patologia , Colo do Útero/cirurgia , Feminino , Seguimentos , Humanos , Técnicas Imunoenzimáticas , Estadiamento de Neoplasias , Prognóstico , Proteômica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/cirurgia , Displasia do Colo do Útero/patologia , Displasia do Colo do Útero/cirurgiaRESUMO
Trefoil factors, which bear a unique 3-loop trefoil domain, are a family of small secretory protease-resistant peptides (7-12 kDa) discovered in the 1980s. Trefoil factor 2 (TFF2) is a unique member of trefoil factors family that plays important roles in gastrointestinal mucosal defense and repair. However, few studies have characterized the miRNA expression patterns in TFF2 knock-out mice. In this study, we investigated the regulatory role of miRNAs in TFF2 knock-out mice. Whole miRNome profiling for TFF2 knock-out mice and wild-type mice were downloaded from the Gene Expression Omnibus database. A total of 14 differentially expressed miRNAs were identified using the limma package. Target genes for 2 differentially expressed miRNAs were retrieved from 2 databases. After mapping these target genes into STRING, an interaction network was constructed. Gene Ontology analysis suggested that the differentially expressed miRNAs are involved in cyclic AMP metabolism and the growth process. Additionally, dysregulated miRNAs target pathways of transforming growth factor-beta signaling pathway and cytokine-cytokine receptor interaction. Our results suggest that miRNAs may play important regulatory roles in processes involving TFF2, particularly in the regulation of signal transduction pathways. However, further validation of our results is needed.
Assuntos
Biologia Computacional , Perfilação da Expressão Gênica , MicroRNAs/genética , Mucinas/genética , Proteínas Musculares/genética , Peptídeos/genética , Transcriptoma , Animais , Regulação da Expressão Gênica , Redes Reguladoras de Genes , Camundongos , Camundongos Knockout , Interferência de RNA , RNA Mensageiro/genética , Fator Trefoil-2Assuntos
Transplante de Rim/fisiologia , Soluções para Preservação de Órgãos , Preservação de Órgãos/métodos , Circulação Renal , Adenosina , Alopurinol , Animais , Cães , Feminino , Glutationa , Sobrevivência de Enxerto , Insulina , Transplante de Rim/imunologia , Transplante de Rim/patologia , Masculino , Microcirculação/fisiologia , Rafinose , Reperfusão , Soluções , Transplante AutólogoAssuntos
Antibacterianos/administração & dosagem , Imunossupressores/administração & dosagem , Transplante de Rim/imunologia , Transplante de Fígado/imunologia , Animais , Cães , Feminino , Sobrevivência de Enxerto/efeitos dos fármacos , Imunidade Celular/efeitos dos fármacos , Injeções Intramusculares , Teste de Cultura Mista de Linfócitos , TacrolimoRESUMO
A technique of bile diversion by tube choledochoureterostomy has been devised for the purpose of studying the role of bile in the intestinal absorption of drugs. This method was used in six dogs. No technical difficulties or major complications developed, as are inevitable with alternative methods, including external fistula.