Microarray-analysis of primary lung fibroblasts cell lines (established from 3-4weeks old mice)

Purpose: The goal of this analysis is to compare gene expression profiling of activated-fibroblasts lines (expressing K-RasG12D) to non-activated-fibroblast expressing WT K-Ras. Methods: Fibroblasts lines were obtained after lung-tissue digestion with collagenase. Cells were homogenized in TRIzol reagent and total RNA was extracted according to manufacturer’s protocol. Hybridization of RNA was performed using Affymetrix Mouse-Gene 1.0ST platform by Dana Farber Microarray Core Facility and data analysis was performed using dChip software. Gene expression analysis was conducted using Transcriptome Analysis Console Software (TAC) using comparing method (SKY vs WT). Results: Gene expression analysis revealed deregulation of several canonical genes involved in different signaling pathways, as Wnt, Notch and Hedgehog signaling pathways. Deregulations of ECM remodeling and focal adhesion expression were reported as well. In addition the secreted molecule Hgf was increased in activated fibroblasts compared to WT line. Examination of 2 different mRNA profiles of 3-4weeks old wild type (WT) and SKY (K-RasG12D) lung fibroblasts cell lines