Gene expression profile of human peritoneal mesothelial cells (HPMC) treated with TGF-b1

Long-term peritoneal dialysis is associated with progressive fibrosis of the peritoneum. Epithelial-mesenchymal transition (EMT) of mesothelial cells is an important mechanism involved in peritoneal fibrosis, and TGF-b1 is considered central in this process. We conducted network-based integrated analysis of transcriptomic data to systemically characterize the molecular signature of TGF-b1-stimulated human peritoneal mesothelial cells (HPMCs). Total RNA was extracted from HPMCs cultured under 1 ng/mL TGF-b1 for 6 and 24 h using Trizol (Invitrogen) according to standard methods. For microarray analysis, 0.5 mg of total RNA was used to make biotin-labeled cRNA using the Ambion Illumina cRNA amplification and labeling kit (Ambion, Austin, TX, USA) . cRNA was then column purified and the quality verified by Agilent 2100 Bioanalyzer (Palo Alto, CA, USA) prior to hybridization. Biotin-labeled cRNA was labeled with Cy3 (Amersham, Piscataway, NJ, USA) and hybridized onto a HumanRef-8 24 K Expression Array Bead Chip (Illumina, San Diego, CA, USA). The arrays were then scanned by an Illumina Bead Station laser scanning imaging system. The data were analyzed using Genespring software from Agilent after normalization.