Gene expression in healthy and lesional psoriatic epidermis from human skin biopsies (laser capture microdissection): set 2

We performed microarray analysis on laser-microdissected epidermis from healthy volunteers and psoriatic patients to identify genes dysregulated in lesional psoriatic epidermis Epidermis and dermis were obtained from human skin biopsies by laser capture microdissection using Arcturus XT (MDS Analytical Technologies) system, followed by RNA isolation with Arcturus Picopure RNA isolation Kit. 10 ng RNA were amplified with ExpressArt Kit and labeled with One-color microarray-based Gene Expression Analysis Low Input Quick Amp WT Labeling (Agilent Technologies) and hybridized with Human Gene Expression Microarray 8x60K chip (Agilent Technologies). Two sets of samples (3 lesional psoriatic and 2 healthy epidermis samples in set 1 and 3 lesional psoriatic and 3 healthy epidermis samples in set 2, or 3 lesional psoriatic and 2 healthy dermis samples in set 1 and 3 lesional psoriatic and 2 healthy dermis samples in set 2) were processed and normalized separately.

本研究针对健康志愿者与银屑病患者经激光显微切割获取的表皮组织开展微阵列分析（microarray analysis），旨在鉴定银屑病病变表皮中失调的基因。研究人员通过搭载Arcturus XT（MDS Analytical Technologies）系统的激光捕获显微切割（Laser Capture Microdissection, LCM）技术，从人类皮肤活检标本中分离得到表皮与真皮组织，随后采用Arcturus Picopure RNA提取试剂盒完成RNA分离。取10 ng RNA，通过ExpressArt试剂盒进行扩增，并利用安捷伦科技（Agilent Technologies）的单通道微阵列基因表达分析低起始量快速扩增WT标记试剂盒（One-color microarray-based Gene Expression Analysis Low Input Quick Amp WT Labeling）完成标记，之后与安捷伦科技的人类基因表达微阵列8x60K芯片（Human Gene Expression Microarray 8x60K chip）进行杂交。本研究设置两套样本，包含两种实验分组方案：方案一为第一套样本含3份银屑病病变表皮样本与2份健康表皮样本，第二套样本含3份银屑病病变表皮样本与3份健康表皮样本；方案二为第一套样本含3份银屑病病变真皮样本与2份健康真皮样本，第二套样本含3份银屑病病变真皮样本与2份健康真皮样本，两套样本均分别进行处理与归一化分析。