title: Similar NF-κB gene signatures in TNF-α treated human endothelial cells and breast tumor biopsies.
abstract: BACKGROUND
  Endothelial dysfunction has been implicated in the pathogenesis of diverse pathologies ranging from vascular and immune diseases to cancer. TNF-α is one of the mediators of endothelial dysfunction through the activation of transcription factors, including NF-κB. While HUVEC (macrovascular cells) have been largely used in the past, here, we documented an NF-κB gene signature in TNFα-stimulated microvascular endothelial cells HMEC often used in tumor angiogenesis studies.
  
  
  RESULTS
  We measured mRNA expression of 55 NF-κB related genes using quantitative RT-PCR in HUVEC and HMEC. Our study identified twenty genes markedly up-regulated in response to TNFα, including adhesion molecules, cytokines, chemokines, and apoptosis regulators, some of them being identified as TNF-α-inducible genes for the first time in endothelial cells (two apoptosis regulators, TNFAIP3 and TNFRSF10B/Trail R2 (DR5), the chemokines GM-CSF/CSF2 and MCF/CSF1, and CD40 and TNF-α itself, as well as NF-κB components (RELB, NFKB1 or 50/p105 and NFKB2 or p52/p100). For eight genes, the fold induction was much higher in HMEC, as compared to HUVEC. Most importantly, our study described for the first time a connection between NF-κB activation and the induction of most, if not all, of these genes in HMEC as evaluated by pharmacological inhibition and RelA expression knock-down by RNA interference. Moreover, since TNF-α is highly expressed in tumors, we further applied the NF-κB gene signature documented in TNFα-stimulated endothelial cells to human breast tumors. We found a significant positive correlation between TNF and the majority (85 %) of the identified endothelial TNF-induced genes in a well-defined series of 96 (48 ERα positive and 48 ERα negative) breast tumors.
  
  
  CONCLUSIONS
  Taken together these data suggest the potential use of this NF-κB gene signature in analyzing the role of TNF-α in the endothelial dysfunction, as well as in breast tumors independently of the presence of ERα.
authors.affiliation: INSERM U965, Paris, France. martine.applanat@inserm.fr
authors.forename: Martine
authors.initials: M
authors.lastname: Perrot-Applanat
authors.forename: Sophie
authors.initials: S
authors.lastname: Vacher
authors.forename: Aurore
authors.initials: A
authors.lastname: Toullec
authors.forename: Irma
authors.initials: I
authors.lastname: Pelaez
authors.forename: Guillaume
authors.initials: G
authors.lastname: Velasco
authors.forename: Françoise
authors.initials: F
authors.lastname: Cormier
authors.forename: Hanan El Sheikh
authors.initials: Hel S
authors.lastname: Saad
authors.forename: Rosette
authors.initials: R
authors.lastname: Lidereau
authors.forename: Véronique
authors.initials: V
authors.lastname: Baud
authors.forename: Ivan
authors.initials: I
authors.lastname: Bièche
mesh_terms: D000368:Aged; D001706:Biopsy; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D055806:Microvessels / Q000473:pathology; D008875:Middle Aged; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*
chemical_list: C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D016328:NF-kappa B; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha