title: STAT1-dependent expression of energy metabolic pathways links tumour growth and radioresistance to the Warburg effect.
abstract: BACKGROUND
  The Signal Transducer and Activator of Transcription 1 (STAT1) has traditionally been regarded as a transmitter of interferon signaling and a pro-apoptotic tumour suppressor. Recent data have identified new functions of STAT1 associated with tumourigenesis and resistance to genotoxic stress, including ionizing radiation (IR) and chemotherapy. To investigate the mechanisms contributing to the tumourigenic functions of STAT1, we performed a combined transcriptomic-proteomic expressional analysis and found that STAT1 is associated with regulation of energy metabolism with potential implication in the Warburg effect.
  
  
  METHODS
  We generated a stable knockdown of STAT1 in the SCC61 human squamous cell carcinoma cell line, established tumour xenografts in athymic mice, and compared transcriptomic and proteomic profiles of STAT1 wild-type (WT) and knockdown (KD) untreated or irradiated (IR) tumours. Transcriptional profiling was based on Affymetrix Human GeneChip(R) Gene 1.0 ST microarrays. Proteomes were determined from the tandem mass spectrometry (MS/MS) data by searching against the human subset of the UniProt database. Data were analysed using Significance Analysis of Microarrays for ribonucleic acid and Visualize software for proteins. Functional analysis was performed with Ingenuity Pathway Analysis with statistical significance measured by Fisher's exact test.
  
  
  RESULTS
  Knockdown of STAT1 led to significant growth suppression in untreated tumours and radio sensitization of irradiated tumours. These changes were accompanied by alterations in the expression of genes and proteins of glycolysis/gluconeogenesis (GG), the citrate cycle (CC) and oxidative phosphorylation (OP). Of these pathways, GG had the most concordant changes in gene and protein expression and demonstrated a STAT1-dependent expression of genes and proteins consistent with tumour-specific glycolysis. In addition, IR drastically suppressed the GG pathway in STAT1 KD tumours without significant change in STAT1 WT tumours.
  
  
  CONCLUSIONS
  Our results identify a previously uncharacterized function of STAT1 in tumours: expressional regulation of genes encoding proteins involved in glycolysis, the citrate cycle and mitochondrial oxidative phosphorylation, with predominant regulation of glycolytic genes. STAT1-dependent expressional regulation of glycolysis suggests a potential role for STAT1 as a transcriptional modulator of genes responsible for the Warburg effect.
authors.affiliation: Department of Radiation and Cellular Oncology, The University of Chicago, Chicago, IL 60637, USA. spitroda@uchicago.edu
authors.forename: Sean P
authors.initials: SP
authors.lastname: Pitroda
authors.forename: Bassam T
authors.initials: BT
authors.lastname: Wakim
authors.forename: Ravi F
authors.initials: RF
authors.lastname: Sood
authors.forename: Mara G
authors.initials: MG
authors.lastname: Beveridge
authors.forename: Michael A
authors.initials: MA
authors.lastname: Beckett
authors.forename: Dhara M
authors.initials: DM
authors.lastname: MacDermed
authors.forename: Ralph R
authors.initials: RR
authors.lastname: Weichselbaum
authors.forename: Nikolai N
authors.initials: NN
authors.lastname: Khodarev
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004734:Energy Metabolism; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D006801:Humans; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy / Q000532:radiotherapy*; D020411:Oligonucleotide Array Sequence Analysis; D020543:Proteome / Q000032:analysis; D011836:Radiation Tolerance; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D020543:Proteome; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human