title: β1-Integrin via NF-κB signaling is essential for acquisition of invasiveness in a model of radiation treated in situ breast cancer.
abstract: BACKGROUND
  Ductal carcinoma in situ (DCIS) is characterized by non-invasive cancerous cell growth within the breast ducts. Although radiotherapy is commonly used in the treatment of DCIS, the effect and molecular mechanism of ionizing radiation (IR) on DCIS are not well understood, and invasive recurrence following radiotherapy remains a significant clinical problem. This study investigated the effects of IR on a clinically relevant model of Akt-driven DCIS and identified possible molecular mechanisms underlying invasive progression in surviving cells.
  
  
  METHODS
  We measured the level of phosphorylated-Akt (p-Akt) in a cohort of human DCIS specimens by immunohistochemistry (IHC) and correlated it with recurrence risk. To model human DCIS, we used Akt overexpressing human mammary epithelial cells (MCF10A-Akt) which, in three-dimensional laminin-rich extracellular matrix (lrECM) and in vivo, form organotypic DCIS-like lesions with lumina expanded by pleiomorphic cells contained within an intact basement membrane. In a population of cells that survived significant IR doses in three-dimensional lrECM, a malignant phenotype emerged creating a model for invasive recurrence.
  
  
  RESULTS
  P-Akt was up-regulated in clinical DCIS specimens and was associated with recurrent disease. MCF10A-Akt cells that formed DCIS-like structures in three-dimensional lrECM showed significant apoptosis after IR, preferentially in the luminal compartment. Strikingly, when cells that survived IR were repropagated in three-dimensional lrECM, a malignant phenotype emerged, characterized by invasive activity, up-regulation of fibronectin, α5β1-integrin, matrix metalloproteinase-9 (MMP-9) and loss of E-cadherin. In addition, IR induced nuclear translocation and binding of nuclear factor-kappa B (NF-κB) to the β1-integrin promoter region, associated with up-regulation of α5β1-integrins. Inhibition of NF-κB or β1-integrin signaling abrogated emergence of the invasive activity.
  
  
  CONCLUSIONS
  P-Akt is up-regulated in some human DCIS lesions and is possibly associated with recurrence. MCF10A-Akt cells form organotypic DCIS-like lesions in three-dimensional lrECM and in vivo, and are a plausible model for some forms of human DCIS. A population of Akt-driven DCIS-like spheroids that survive IR progresses to an invasive phenotype in three-dimensional lrECM mediated by β1-integrin and NF-κB signaling.
authors.forename: Jin-Min
authors.initials: JM
authors.lastname: Nam
authors.forename: Kazi M
authors.initials: KM
authors.lastname: Ahmed
authors.forename: Sylvain
authors.initials: S
authors.lastname: Costes
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Onodera
authors.forename: Adam B
authors.initials: AB
authors.lastname: Olshen
authors.forename: Kanako C
authors.initials: KC
authors.lastname: Hatanaka
authors.forename: Rumiko
authors.initials: R
authors.lastname: Kinoshita
authors.forename: Masayori
authors.initials: M
authors.lastname: Ishikawa
authors.forename: Hisataka
authors.initials: H
authors.lastname: Sabe
authors.forename: Hiroki
authors.initials: H
authors.lastname: Shirato
authors.forename: Catherine C
authors.initials: CC
authors.lastname: Park
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000528:radiation effects; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology* / Q000532:radiotherapy; D002285:Carcinoma, Intraductal, Noninfiltrating / Q000378:metabolism / Q000473:pathology / Q000532:radiotherapy; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D004789:Enzyme Activation; D005260:Female; D064593:Heterografts; D006801:Humans; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009364:Neoplasm Recurrence, Local; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism; D011839:Radiation, Ionizing; D015398:Signal Transduction; D018874:Spheroids, Cellular; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
chemical_list: D019012:Integrin beta1; D016328:NF-kappa B; D051057:Proto-Oncogene Proteins c-akt