title: The silencing of the SWI/SNF subunit and anticancer gene BRM in Rhabdoid tumors.
abstract: Rhabdoid sarcomas are highly malignant tumors that usually occur in young children. A key to the genesis of this tumor is the mutational loss of the BAF47 gene as well as the widespread epigenetic suppression of other key anticancer genes. The BRM gene is one such epigenetically silenced gene in Rhabdoid tumors. This gene codes for an ATPase catalytic subunit that shifts histones and opens the chromatin. We show that BRM is an epigenetically silenced gene in 10/11 Rhabdoid cell lines and in 70% of Rhabdoid tumors. Moreover, BRM can be induced by BAF47 re-expression and by Flavopiridol. By selective shRNAi knockdown of BRM, we show that BRM re-expression is necessary for growth inhibition by BAF47 re-expression or Flavopiridol application. Similar to lung cancer cell lines, we found that HDAC3, HDAC9, MEF2D and GATA3 controlled BRM silencing and that HDAC9 was overexpressed in Rhabdoid cancer cell lines. In primary BRM-deficient Rhabdoid tumors, HDAC9 was also found to be highly overexpressed. Two insertional BRM promoter polymorphisms contribute to BRM silencing, but only the -1321 polymorphism correlated with BRM silencing in Rhabdoid cell lines. To determine how these polymorphisms were tied to BRM silencing, we conducted ChIP assays and found that both HDAC9 and MEF2D bound to the BRM promoter at or near these polymorphic sites. Using BRM promoter swap experiments, we indirectly showed that both HDAC9 and MEF2D bound to these polymorphic sites. Together, these data show that the mechanism of BRM silencing contributes to the pathogenesis of Rhabdoid tumors and appears to be conserved among tumor types.
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, University of Florida, Florida, USA.
authors.forename: Bhaskar
authors.initials: B
authors.lastname: Kahali
authors.forename: Jinlong
authors.initials: J
authors.lastname: Yu
authors.forename: Stefanie B
authors.initials: SB
authors.lastname: Marquez
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Thompson
authors.forename: Shermi Y
authors.initials: SY
authors.lastname: Liang
authors.forename: Li
authors.initials: L
authors.lastname: Lu
authors.forename: David
authors.initials: D
authors.lastname: Reisman
mesh_terms: D015153:Blotting, Western; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics*; D055785:Gene Knockdown Techniques; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018335:Rhabdoid Tumor / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D014162:Transfection
chemical_list: D002868:Chromosomal Proteins, Non-Histone; C084011:SMARCA2 protein, human; C445179:SWI-SNF-B chromatin-remodeling complex; D014157:Transcription Factors