title: RNA-Seq and Network Analysis Revealed Interacting Pathways in TGF-β-Treated Lung Cancer Cell Lines.
abstract: Whole transcriptome shotgun sequencing (RNA-Seq) is a useful tool for analyzing the transcriptome of a biological sample. With appropriate statistical and bioinformatic processing, this platform is capable of identifying significant differences in gene expression within the transcriptome and permits pathway and network analyses to determine how these genes interact biologically. In this study, we examined gene expression in two lung adenocarcinoma cell lines (H358 and A459) that were treated with transforming growth factor-β (TGF-β) as a model for induction of the epithelial-to-mesenchymal transition (EMT), commonly associated with disease progression. We performed this study in order to illustrate a workflow for identifying interesting genes and processes that are regulated early in EMT and to determine their gene pathway/network relationships and regulation. With this, we identified 137 upregulated and 32 downregulated genes common to both cell lines after TGF-β treatment that represent components of multiple canonical pathways and biological networks associated with the induction of EMT. These findings were also verified against reposited Affymetrix U133a expression profiles from multiple trials examining metastatic progression in patient cohorts (n = 731 total) to further establish the clinical relevance and translational significance of the model system. Together, these findings help validate the relevance of the TGF-β model for the study of EMT and provide new insights into early events in EMT.
authors.affiliation: Department of Internal Medicine, Rush University Medical Center, Chicago, IL, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Biochemistry, Rush University Medical Center, Chicago, IL, USA.
authors.forename: Omid
authors.initials: O
authors.lastname: Rouhi
authors.affiliation: Department of Internal Medicine, Rush University Medical Center, Chicago, IL, USA.
authors.forename: Hankui
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Internal Medicine, Rush University Medical Center, Chicago, IL, USA.
authors.forename: Rolando
authors.initials: R
authors.lastname: Ramirez
authors.affiliation: Department of Biochemistry, Rush University Medical Center, Chicago, IL, USA. ; Department of Pathology, Rush University Medical Center, Chicago, IL, USA.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Borgia
authors.affiliation: Department of Internal Medicine, Rush University Medical Center, Chicago, IL, USA. ; Department of Biochemistry, Rush University Medical Center, Chicago, IL, USA.
authors.forename: Youping
authors.initials: Y
authors.lastname: Deng
keywords: RNA-seq; TGF treatment; gene expression; gene pathways; lung cancer; network analysis