cDNA macroarray analysis of genes in renal epithelial cells exposed to calcium oxalate crystals
cDNA macroarray analysis of genes in renal epithelial cells exposed to calcium oxalate crystals
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DOI：
10.1007/s00240-008-0164-2
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发表时间：
2009-02-01
影响因子：
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通讯作者：
Suzuki, Koji
中科院分区：
文献类型：
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作者：
Miyazawa, Katsuhito;Aihara, Kinue;Suzuki, Koji
肾结石的形成是一个复杂的过程，许多基因参与这一级联反应。一水草酸钙（COM）晶体是肾结石中最常见的晶体，肾上皮细胞的结合和内化可能是导致肾结石形成的关键步骤。暴露于 COM 晶体会改变多种基因的表达，但之前对基因表达的研究普遍有限。为了更详细地了解基因表达，我们使用 cDNA 宏阵列检查了暴露于 COM 晶体的肾上皮细胞中的基因表达谱。 NRK-52E 细胞暴露于 COM 晶体 60 和 120 分钟。分离Poly (A)(+) RNA并转化为P-32标记的第一链cDNA，然后将cDNA探针与膜杂交。扫描杂交图像并对信号强度进行量化。使用全局总和归一化方法分析了 1,176 个基因的 mRNA 表达。暴露于 COM 晶体会改变之前报道的一些基因的表达。此外，还鉴定出了新的基因。超过 20 个基因被发现至少受到双重调控。我们对暴露于 COM 晶体的肾上皮细胞的基因表达进行了大规模分析，并鉴定了差异调节的基因。 cDNA 宏阵列是评估尿石症研究中基因表达的有用工具。
Kidney stone formation is a complex process, and numerous genes participate in this cascade. The binding and internalization of calcium oxalate monohydrate (COM) crystals, the most common crystal in renal stones by renal epithelial cells may be a critical step leading to kidney stone formation. Exposure to COM crystals alters the expression of various genes, but previous studies on gene expression have generally been limited. To obtain more detailed insight into gene expression, we examined gene expression profiles in renal epithelial cells exposed to COM crystals using cDNA macroarray. NRK-52E cells were exposed to COM crystals for 60 and 120 min. Poly (A)(+) RNA was isolated and converted into P-32-labeled first-strand cDNA, then the cDNA probe was hybridized to the membrane. Hybridization images were scanned and the signal intensities were quantified. Expression of mRNA of 1,176 genes was analyzed with global sum normalization methods. Exposure to COM crystals altered the expression of some of the genes reported previously. Furthermore, novel genes were also identified. Over 20 genes were found to be regulated at least twofold. We performed a large-scale analysis of gene expression in renal epithelial cells exposed to COM crystals, and identified the genes differentially regulated. cDNA macroarray is a useful tool for evaluating gene expression in urolithiasis research.