Microarray study of human adrenal zona glomerulosa (ZG), zona fasciculata (ZF) and aldosterone-producing adenomas (APA). Homo sapiens

Learn about the transcriptome profiling of zona glomerulosa (ZG), zona fasciculata (ZF) and aldosterone-producing adenomas (APA) in human adrenals Overall design: 21 pairs of zona fasciculata (ZF) and zona glomerulosa (ZG), and 14 paired aldosterone-producing adenomas (APAs) from 14 Conn’s syndrome patients and 7 phaeochromocytoma patients were assayed on the Affymetrix Human Genome U133 Plus 2.0 Array. Laser capture microdissection was used to acquire samples of ZF, ZG and APA as previously described (Azizan EA, et al. J Clin Endocrinol Metab. 2012;97:E819-E829). For differentiation of ZG from ZF, sections were stained with cresyl violet using the LCM Staining Kit (AM1935, Ambion, USA). Data processing and analysis was performed using AffymetrixGeneChip Command Console Software and PartekGenomicSuite 6.5 (Partek Inc., St. Louis, MO). Gene expressions were portrayed as the summarized log-signal of the Robust Multichip Average (RMA) with quantilenormalisation and median polish for probe set summarisation. Validation by qPCR was performed on genes >10 fold up-regulated in zona glomerulosa (compared to zona fasciculata) and >10 fold up-regulated in aldosterone-producing adenomas (compared to zona glomerulosa).

本数据集旨在解析人类肾上腺球状带（zona glomerulosa, ZG）、束状带（zona fasciculata, ZF）以及醛固酮分泌性腺瘤（aldosterone-producing adenomas, APA）的转录组表达谱。实验设计：共收集21对束状带（ZF）与球状带（ZG）配对样本，以及14例配对醛固酮分泌性腺瘤（APA）样本，上述样本分别来自14名Conn综合征患者与7名嗜铬细胞瘤患者，均采用Affymetrix人类基因组U133 Plus 2.0基因芯片完成表达检测。样本获取采用激光捕获显微切割（Laser capture microdissection, LCM）技术，具体操作参照既往研究（Azizan EA等，J Clin Endocrinol Metab. 2012;97:E819-E829）。为区分球状带与束状带组织，使用LCM染色试剂盒（AM1935，Ambion公司，美国）的焦油紫对组织切片进行染色。数据处理与分析采用Affymetrix GeneChip 命令控制台软件及Partek GenomicSuite 6.5（Partek公司，美国密苏里州圣路易斯市）完成。基因表达量以稳健多数组平均（Robust Multichip Average, RMA）的汇总对数信号值表示，该分析通过分位数归一化与中位数抛光法完成探针组的汇总计算。通过定量PCR（qPCR）对两类差异基因进行验证：即在球状带中较束状带上调10倍以上的基因，以及在醛固酮分泌性腺瘤中较球状带上调10倍以上的基因。