Expression data from human liver cell line induced by PCB153

Exposure to Polychlorobiphenyls (PCBs) is known to cause serious health effects in human but the gene expression profiles leading to development of differnet diseases is not fully understood. The knowledge of global gene expression will help us to devlop early diagnostic biomarkers for PCB induced health effects. We used microarrays to detail the global gene expression profile underlying the effects of PCB153 on human liver cells leading to identification of distinct classes of up-regulated and down-regulated genes and cellular processes. Keywords: time course The HepG2 cells were grown for 48 hours in DMEM supplemented with 10% FBS and 1X Pennicillin-Streptromycin and treated with PCB153 for different times. Trizol extraction of total RNA was performed according to the manufacturer's instructions. Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 6 microgram of total RNA (Expression Analysis Technical Manual, 2001, Affymetrix). Following fragmentation, 10 microgram of cRNA were hybridized for 16 hr at 45C on GeneChip Human Genome Array (HGU133 plus 2.0). GeneChips were washed and stained in the Affymetrix Fluidics Station 450 and scanned using the Affymetrix GeneArray Scanner 3000. The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitrarily set to 150.

已知多氯联苯（Polychlorobiphenyls, PCBs）暴露可对人体造成严重健康损害，但目前尚未完全明确其诱发各类疾病的基因表达调控机制。阐明全局基因表达谱将有助于开发多氯联苯暴露所致健康损害的早期诊断生物标志物。本研究采用基因芯片（microarrays）解析PCB153作用于人肝细胞后的全局基因表达谱，以甄别显著上调、下调的基因类别及相关细胞生物学过程。关键词：时间序列

将HepG2细胞置于添加10%胎牛血清（Fetal Bovine Serum, FBS）与1×青霉素-链霉素的达尔伯克改良伊格尔培养基（DMEM）中培养48小时，随后用PCB153处理不同时长。按照试剂制造商的操作说明，采用Trizol法提取总RNA。取6μg总RNA，依照Affymetrix标准实验流程（《表达分析技术手册》，2001年，Affymetrix）制备生物素标记的cRNA。经片段化处理后，取10μg cRNA在45℃下与GeneChip人类基因组阵列（HGU133 plus 2.0）杂交16小时。将基因芯片置于Affymetrix Fluidics Station 450中完成洗涤与染色，随后使用Affymetrix GeneArray Scanner 3000进行扫描。采用Microarray Suite 5.0（MAS 5.0）分析数据，使用Affymetrix默认分析参数，并以全局缩放作为标准化方法；将各芯片的截尾均值靶标强度统一设置为150。