RESUMO
The importance of stromal cells and the factors that they expressed during cancer initiation and progression have been highlighted by recent literature. To identify the stromal proteins involved in nasopharyngeal carcinoma (NPC) carcinogenesis, we assessed differences in protein expression of the stroma from NPC and normal nasopharyngeal epithelium tissues (NNET) using a quantitative proteomic approach combined with laser capture microdissection (LCM). LCM was performed to purify stromal cells from the NPC and NNET, respectively. The differential proteins between the pooled microdissected tumor and normal stroma were analyzed by two-dimensional difference gel electrophoresis (2D-DIGE) combined with mass spectrometry (MS). Twenty differential proteins were identified, and the expression and location of two differential proteins (L-plastin and S100A9) were further confirmed by Western blotting and immunohistochemical analysis. Our results will be helpful to study the role of stroma in the NPC carcinogenesis, as well as discover the interaction between NPC cells and their surrounding microenvironment.
Assuntos
Calgranulina B/análise , Tecido Conjuntivo/química , Glicoproteínas de Membrana/análise , Proteínas dos Microfilamentos/análise , Neoplasias Nasofaríngeas/química , Proteínas de Neoplasias/análise , Proteômica/métodos , Eletroforese em Gel Bidimensional , Epitélio/química , Feminino , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Neoplasias Nasofaríngeas/patologia , Nasofaringe/química , Nasofaringe/citologiaRESUMO
Metabolomics, as a promising and powerful approach, refers to comprehensive assessment and identification of small molecule endogenous metabolites in a biological system which is capable of further understanding the mechanisms of diseases for early diagnosis, effective treatment and prognosis. Acute kidney injury (AKI) induced by contrast is a serious complication in patients undergoing administration of iodinated contrast media. It is becoming a major health concern in clinic, however, the molecular mechanisms of contrast-induced acute kidney injury (CI-AKI) have not been well characterized. In this study, we used serum metabolomics based on liquid chromatography-mass spectrometry (LC-MS) combined with pattern recognition to explore and characterize potential metabolites and metabolic pathway in an experimental model for CI-AKI. Seventeen differentiating metabolites in the serum were identified involving the pivotal metabolic pathways related to tryptophan metabolism, glycerophospholipid metabolism, steroid hormone biosynthesis, pyrimidine metabolism, sphingolipid metabolism, aminoacyl-tRNA biosynthesis. Our study provides novel insight into pathophysiologic mechanisms of AKI by changing biomarkers and pathways.
RESUMO
The stroma surrounding cancer cell population is increasingly recognized as playing an important role in cancer proliferation, invasion, and metastasis. To identify the stromal proteins involved in nasopharyngeal carcinoma (NPC) carcinogenesis, differences in protein expression of the stroma from NPC and normal nasopharyngeal epithelium tissues (NNET) were assessed using a comparative proteomic approach combined with laser capture microdissection (LCM). LCM was performed to purify stromal cells from NPC and NNET, respectively. Proteins between the pooled microdissected tumor and normal stroma were separated by two-dimensional electrophoresis (2-DE) and differential proteins were identified by mass spectrometry (MS). Sixty differential proteins between normal stroma (NS) and tumor stroma (TS) were identified, and the expression of CapG protein was further confirmed by western blotting and immunohistochemical analysis. Our results will be helpful to study the role of stroma in the NPC carcinogenesis and may provide helpful clues for pathogenesis, early diagnosis, and progression of NPC.
Assuntos
Carcinoma de Células Escamosas/química , Carcinoma de Células Escamosas/patologia , Neoplasias Nasofaríngeas/química , Neoplasias Nasofaríngeas/patologia , Nasofaringe/química , Nasofaringe/citologia , Proteoma/análise , Adulto , Idoso , Sequência de Aminoácidos , Carcinoma de Células Escamosas/metabolismo , Eletroforese em Gel Bidimensional , Epitélio/química , Epitélio/metabolismo , Feminino , Humanos , Masculino , Espectrometria de Massas , Microdissecção , Proteínas dos Microfilamentos/análise , Proteínas dos Microfilamentos/biossíntese , Pessoa de Meia-Idade , Dados de Sequência Molecular , Neoplasias Nasofaríngeas/metabolismo , Nasofaringe/metabolismo , Proteínas de Neoplasias/análise , Proteínas de Neoplasias/biossíntese , Proteínas Nucleares/análise , Proteínas Nucleares/biossíntese , Proteoma/biossíntese , Células Estromais/química , Células Estromais/metabolismoRESUMO
In this study, we aim to screen metastasis-related proteins in human lung squamous carcinoma (LSC) using laser capture microdissection and a proteomic approach. Twenty two differential proteins were identified from pooled microdissected primary LSC and matched lymph node (LN) metastatic tissues. Expression of the differential protein 14-3-3 sigma was determined by Western blotting and immunohistochemistry. In cell invasion assay, down-regulated 14-3-3 sigma by siRNA increased in vitro invasive ability of HTB-182 and A549 cells, up-regulation of 14-3-3 sigma by pcDNA3.0/14-3-3 sigma decreased in vitro invasive ability of HTB-182 and A549 cells. The data suggest that 14-3-3 sigma is a potential LN metastasis-related protein in LSC, and its dysregulation might play an important role in the LN metastatic process of LSC.