title: Interferon-γ-induced activation of Signal Transducer and Activator of Transcription 1 (STAT1) up-regulates the tumor suppressing microRNA-29 family in melanoma cells.
abstract: BACKGROUND
  The type-II-cytokine IFN-γ is a pivotal player in innate immune responses but also assumes functions in controlling tumor cell growth by orchestrating cellular responses against neoplastic cells. The role of IFN-γ in melanoma is not fully understood: it is a well-known growth inhibitor of melanoma cells in vitro. On the other hand, IFN-γ may also facilitate melanoma progression. While interferon-regulated genes encoding proteins have been intensively studied since decades, the contribution of miRNAs to effects mediated by interferons is an emerging area of research.We recently described a distinct and dynamic regulation of a whole panel of microRNAs (miRNAs) after  IFN-γ-stimulation. The aim of this study was to analyze the transcriptional regulation of miR-29 family members  in detail, identify potential interesting target genes and thus further elucidate a potential signaling pathway  IFN-γ → Jak→ P-STAT1 → miR-29 → miR-29 target genes and its implication for melanoma growth.
  
  
  RESULTS
  Here we show that IFN-γ induces STAT1-dependently a profound up-regulation of the miR-29 primary cluster pri-29a~b-1 in melanoma cell lines. Furthermore, expression levels of pri-29a~b-1 and mature miR-29a and miR-29b were elevated while the pri-29b-2~c cluster was almost undetectable. We observed an inverse correlation between miR-29a/b expression and the proliferation rate of various melanoma cell lines. This finding could be corroborated in cells transfected with either miR-29 mimics or inhibitors. The IFN-γ-induced G1-arrest of melanoma cells involves down-regulation of CDK6, which we proved to be a direct target of miR-29 in these cells. Compared to nevi and normal skin, and metastatic melanoma samples, miR-29a and miR-29b levels were found strikingly elevated in certain patient samples derived from primary melanoma.
  
  
  CONCLUSIONS
  Our findings reveal that the miR-29a/b1 cluster is to be included in the group of IFN- and STAT-regulated genes. The up-regulated miR-29 family members may act as effectors of cytokine signalling in melanoma and other cancer cells as well as in the immune system.
authors.affiliation: Signal Transduction Laboratory, University of Luxembourg, 162A Avenue de la Faïencerie, Luxembourg, L-1511, Luxembourg. Stephanie.Kreis@uni.lu.
authors.forename: Martina J
authors.initials: MJ
authors.lastname: Schmitt
authors.forename: Demetra
authors.initials: D
authors.lastname: Philippidou
authors.forename: Susanne E
authors.initials: SE
authors.lastname: Reinsbach
authors.forename: Christiane
authors.initials: C
authors.lastname: Margue
authors.forename: Anke
authors.initials: A
authors.lastname: Wienecke-Baldacchino
authors.forename: Dorothee
authors.initials: D
authors.lastname: Nashan
authors.forename: Iris
authors.initials: I
authors.lastname: Behrmann
authors.forename: Stephanie
authors.initials: S
authors.lastname: Kreis