Differential gene expression in astrocytes from human normal and glaucomatous optic nerve head analyzed by cDNA microarray
Differential gene expression in astrocytes from human normal and glaucomatous optic nerve head analyzed by cDNA microarray
复制标题
作者：
Hernandez, MR;Agapova, OA;Aoi, S
cDNA微阵列技术的最新进展使得同时分析数千个基因的表达成为可能。使用这种技术，在人星形胶质细胞的基因表达进行了比较，从脑胶质瘤和正常的视神经头（ONH）培养。150个基因在肿瘤细胞培养物中的表达与正常细胞相比差异超过5倍。这些基因参与许多生物学过程，包括信号转导、细胞粘附和增殖、ECM合成和降解。通过定量RT-PCR进行差异基因表达的确认。蛋白质印迹和免疫组化证明基因产物在细胞培养物或人ONH组织中。增殖，粘附和迁移试验测试差异基因表达所提示的生理反应。我们的研究表明，培养的青光眼ONH星形胶质细胞在培养中保留了许多表型特征，这些表型特征可能与它们在青光眼发病机制中的作用有关，并且通常与CNS中的反应性星形胶质细胞有关。这些数据的潜在应用包括星形胶质细胞功能相关信号通路的识别和表征，类固醇代谢酶在青光眼ONH中作用的研究，以及进一步探索选定的已识别基因在实验动物和体外青光眼模型中的作用。(C)2002 Wiley-Liss，Inc.
Recent advances in cDNA microarray technology have made it possible to analyze expression of several thousand genes at the same time. Using this technique, gene expression in human astrocytes cultured from glaucomatous and normal Optic nerve heads (ONH) was compared. One hundred-fifty genes were differentially expressed more than 5-fold in glaucomatous cell cultures compared with normal. These genes are involved in a number of biological processes, including signal transduction, cell adhesion and proliferation, ECM synthesis, and degradation. Confirmation of differential gene expression was performed by quantitative RT-PCR. Western blots and immunohistochemistry demonstrated gene products in cell cultures or in human ONH tissues. Proliferation, adhesion and migration assays tested physiological responses suggested by differential gene expression. Our study suggests that cultured glaucomatous ONH astrocytes retain in culture many phenotypic characteristics that may be relevant to their role in the pathogenesis of glaucoma and, in general to reactive astrocytes in the CNS. Potential applications of these data include the identification and characterization of signaling pathways involved in astrocyte function, studies of the role of steroid-metabolizing enzymes in the glaucomatous ONH, and further exploration of the role of selected identified genes in experimental animal and in vitro models of glaucoma. (C) 2002 Wiley-Liss, Inc.