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Identification of thioredoxin reductase 1-regulated genes using small interference RNA and cDNA microarray

Thioredoxin reductase 1 (TrxR1) is a cytosolic enzyme that plays a central role in controlling cellular redox homeostasis. TrxR1 can transduce regulatory redox signals through the NADPH-dependent reduction of thioredoxin (Trx), which is able to reduce a broad spectrum of target enzymes and to regulate the activity of several transcription factors (e.g. p53 and NF-£eB). The TrxR1/Trx system is involved in every step of cancer biology, ranging from transformation and progression to invasion, metastasis, and resistance to therapy. TrxR1 was also recently identified as one key enzyme involved in cell death induced by interferon-beta] (IFN-beta])/all-trans retinoic acid (ATRA) anti-cancer treatment. Our study employed small interference RNA (siRNA) and microarray techniques to investigate the effect of TrxR1 silencing on gene expression in HepG2 cells. We also investigated TrxR1-mediated cell response to IFN-beta]/ATRA treatment. We identified TrxR1-dependent genes with functions related to several cellular processes such as apoptosis (SOX4), ubiquitination (Ubiquitin D, F-box protein 25), organization of cytoskeletal/extracellular matrix (Keratin 19, Fibronectin 1), and transport (Cystine/Glutamate transporter). We also investigated the effect of TrxR1 siRNA on the protein profile using surface enhanced laser desorption ionization time-of-flight (SELDI-TOF) technology. Profiles confirmed the significant involvement of TrxR1 in cell response to IFN-beta]/ATRA.