17-β-estradiol (estrogen) is a steroid hormone important to human development;however, high levels of this molecule are associated with increased risk of breast cancer primarily due to estrogen’s ability to bind and...17-β-estradiol (estrogen) is a steroid hormone important to human development;however, high levels of this molecule are associated with increased risk of breast cancer primarily due to estrogen’s ability to bind and activate the estrogen receptor (ER) and initiate gene transcription. Currently, estrogen mechanisms of action are classified as genomic and non-genomic and occur in an ER-dependent and ER-independent manner. In this study, we examine estrogen signaling pathways, by measuring changes in protein expression as a function of time of exposure to estrogen in both ER-positive (MCF-7) and ER-negative (MDA-MB-231) cell lines. Using a robust experimental design utilizing isotopic labeling, two-dimensional LC-MS, and bioinformatics analysis, we report genomic and non-genomic ER regulated estrogen responsive proteins. We find a little over 200 proteins differentially expressed after estrogen treatment. Cell proliferation, transcription, actin filament capping and cell to cell signaling are significantly enriched in the MCF-7 cell line alone. Translational elongation and proteolysis are enriched in both cell lines. Subsets of the proteins presented in this study are for the first time directly associated with estrogen signaling in mammary carcinoma cells. We find that estrogen affected the expression of proteins involved in numerous processes that are related to tumorigenesis such as increased cellular division and invasion in an ER-dependent manner. Moreover, we identified negative regulation of apoptosis as a non-genomic process of estrogen. This study complements gene expression studies and highlights the need for both genomic and proteomic analyses in unraveling the complex mechanisms by which estrogen affects progression of breast cancer.展开更多
细胞培养条件下稳定同位素标记技术(stable isotope labeling by amino acids in cell culture,SILAC)是一种简单的体内标记策略,是基于质谱的定量蛋白质组学技术中的一个有力工具。SILAC技术不会引起蛋白质互作研究中的假阳性结果,使...细胞培养条件下稳定同位素标记技术(stable isotope labeling by amino acids in cell culture,SILAC)是一种简单的体内标记策略,是基于质谱的定量蛋白质组学技术中的一个有力工具。SILAC技术不会引起蛋白质互作研究中的假阳性结果,使细胞信号转导动态变化研究成为可能,并且能够直接测量蛋白质含量。SILAC技术已经应用于一系列的生物学研究中,近来已被成功应用到整体动物水平的定量蛋白质组研究。本文综述了SILAC技术的基本原理、优点和应用进展。展开更多
Most type 2 diabetics are accompanied with deficiency of insulin secretion and hyperglycemia. It has reported that glucose-stimulated insulin secretion of β cell (GSIS)
Brain-derived neurotrophic factor（BDNF） plays an important role in neurodevelopment,synaptic plasticity,learning and memory,and in preventing neurodegeneration.Despite decades of investigations into downstream signa...Brain-derived neurotrophic factor（BDNF） plays an important role in neurodevelopment,synaptic plasticity,learning and memory,and in preventing neurodegeneration.Despite decades of investigations into downstream signaling cascades and changes in cellular processes,the mechanisms of how BDNF reshapes circuits in vivo remain unclear.This informational gap partly arises from the fact that the bulk of studies into the molecular actions of BDNF have been performed in dissociated neuronal cultures,while the majority of studies on synaptic plasticity,learning and memory were performed in acute brain slices or in vivo.A recent study by Bowling-Bhattacharya et al.,measured the proteomic changes in acute adult hippocampal slices following treatment and reported changes in proteins of neuronal and non-neuronal origin that may in concert modulate synaptic release and secretion in the slice.In this paper,we place these findings into the context of existing literature and discuss how they impact our understanding of how BDNF can reshape the brain.展开更多
文摘17-β-estradiol (estrogen) is a steroid hormone important to human development;however, high levels of this molecule are associated with increased risk of breast cancer primarily due to estrogen’s ability to bind and activate the estrogen receptor (ER) and initiate gene transcription. Currently, estrogen mechanisms of action are classified as genomic and non-genomic and occur in an ER-dependent and ER-independent manner. In this study, we examine estrogen signaling pathways, by measuring changes in protein expression as a function of time of exposure to estrogen in both ER-positive (MCF-7) and ER-negative (MDA-MB-231) cell lines. Using a robust experimental design utilizing isotopic labeling, two-dimensional LC-MS, and bioinformatics analysis, we report genomic and non-genomic ER regulated estrogen responsive proteins. We find a little over 200 proteins differentially expressed after estrogen treatment. Cell proliferation, transcription, actin filament capping and cell to cell signaling are significantly enriched in the MCF-7 cell line alone. Translational elongation and proteolysis are enriched in both cell lines. Subsets of the proteins presented in this study are for the first time directly associated with estrogen signaling in mammary carcinoma cells. We find that estrogen affected the expression of proteins involved in numerous processes that are related to tumorigenesis such as increased cellular division and invasion in an ER-dependent manner. Moreover, we identified negative regulation of apoptosis as a non-genomic process of estrogen. This study complements gene expression studies and highlights the need for both genomic and proteomic analyses in unraveling the complex mechanisms by which estrogen affects progression of breast cancer.
文摘细胞培养条件下稳定同位素标记技术(stable isotope labeling by amino acids in cell culture,SILAC)是一种简单的体内标记策略,是基于质谱的定量蛋白质组学技术中的一个有力工具。SILAC技术不会引起蛋白质互作研究中的假阳性结果,使细胞信号转导动态变化研究成为可能,并且能够直接测量蛋白质含量。SILAC技术已经应用于一系列的生物学研究中,近来已被成功应用到整体动物水平的定量蛋白质组研究。本文综述了SILAC技术的基本原理、优点和应用进展。
文摘Most type 2 diabetics are accompanied with deficiency of insulin secretion and hyperglycemia. It has reported that glucose-stimulated insulin secretion of β cell (GSIS)
基金supported by NIH grants NS034007 and NS047384supported by NIH grants NS21072,and HD23315supported by funds from the Department of Biotechnology,Government of India and the Shanta Wadhwani Foundation
文摘Brain-derived neurotrophic factor（BDNF） plays an important role in neurodevelopment,synaptic plasticity,learning and memory,and in preventing neurodegeneration.Despite decades of investigations into downstream signaling cascades and changes in cellular processes,the mechanisms of how BDNF reshapes circuits in vivo remain unclear.This informational gap partly arises from the fact that the bulk of studies into the molecular actions of BDNF have been performed in dissociated neuronal cultures,while the majority of studies on synaptic plasticity,learning and memory were performed in acute brain slices or in vivo.A recent study by Bowling-Bhattacharya et al.,measured the proteomic changes in acute adult hippocampal slices following treatment and reported changes in proteins of neuronal and non-neuronal origin that may in concert modulate synaptic release and secretion in the slice.In this paper,we place these findings into the context of existing literature and discuss how they impact our understanding of how BDNF can reshape the brain.
