title: Identification of novel non-coding RNA-based negative feedback regulating the expression of the oncogenic transcription factor GLI1.
abstract: Non-coding RNAs are a complex class of nucleic acids, with growing evidence supporting regulatory roles in gene expression. Here we identify a non-coding RNA located head-to-head with the gene encoding the Glioma-associated oncogene 1 (GLI1), a transcriptional effector of multiple cancer-associated signaling pathways. The expression of this three-exon GLI1 antisense (GLI1AS) RNA in cancer cells was concordant with GLI1 levels. siRNAs knockdown of GLI1AS up-regulated GLI1 and increased cellular proliferation and tumor growth in a xenograft model system. Conversely, GLI1AS overexpression decreased the levels of GLI1, its target genes PTCH1 and PTCH2, and cellular proliferation. Additionally, we demonstrate that GLI1 knockdown reduced GLI1AS, while GLI1 overexpression increased GLI1AS, supporting the role of GLI1AS as a target gene of the GLI1 transcription factor. Activation of TGFβ and Hedgehog signaling, two known regulators of GLI1 expression, conferred a concordant up-regulation of GLI1 and GLI1AS in cancer cells. Finally, analysis of the mechanism underlying the interplay between GLI1 and GLI1AS indicates that the non-coding RNA elicits a local alteration of chromatin structure by increasing the silencing mark H3K27me3 and decreasing the recruitment of RNA polymerase II to this locus. Taken together, the data demonstrate the existence of a novel non-coding RNA-based negative feedback loop controlling GLI1 levels, thus expanding the repertoire of mechanisms regulating the expression of this oncogenic transcription factor.
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden; Faculty of Natural Sciences and Mathematics & Doctoral Program in Biomedical Sciences, Universidad del Rosario, Bogotá, Colombia.
authors.forename: Victoria E
authors.initials: VE
authors.lastname: Villegas
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Mohammed Ferdous-Ur
authors.initials: MF
authors.lastname: Rahman
authors.affiliation: Schulze Center for Novel Therapeutics, Mayo Clinic, Rochester, MN, USA.
authors.forename: Maite G
authors.initials: MG
authors.lastname: Fernandez-Barrena
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Yumei
authors.initials: Y
authors.lastname: Diao
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Eleni
authors.initials: E
authors.lastname: Liapi
authors.affiliation: Unit of Dermatology and Venereology, Department of Medicine, Karolinska Institutet, Solna, Sweden.
authors.forename: Enikö
authors.initials: E
authors.lastname: Sonkoly
authors.affiliation: Unit of Dermatology and Venereology, Department of Medicine, Karolinska Institutet, Solna, Sweden.
authors.forename: Mona
authors.initials: M
authors.lastname: Ståhle
authors.affiliation: Unit of Dermatology and Venereology, Department of Medicine, Karolinska Institutet, Solna, Sweden.
authors.forename: Andor
authors.initials: A
authors.lastname: Pivarcsi
authors.affiliation: Department of Medical Sciences, University of Turin, Turin, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Annaratone
authors.affiliation: Department of Medical Sciences, University of Turin, Turin, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Sapino
authors.affiliation: Faculty of Natural Sciences and Mathematics & Doctoral Program in Biomedical Sciences, Universidad del Rosario, Bogotá, Colombia.
authors.forename: Sandra
authors.initials: S
authors.lastname: Ramírez Clavijo
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Bürglin
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimokawa
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
authors.forename: Saraswathi
authors.initials: S
authors.lastname: Ramachandran
authors.affiliation: St. Laurent Institute, One Kendall Square, Cambridge, MA, USA.
authors.forename: Philipp
authors.initials: P
authors.lastname: Kapranov
authors.affiliation: Schulze Center for Novel Therapeutics, Mayo Clinic, Rochester, MN, USA.
authors.forename: Martin E
authors.initials: ME
authors.lastname: Fernandez-Zapico
authors.affiliation: Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden. Electronic address: peter.zaphiropoulos@ki.se.
authors.forename: Peter G
authors.initials: PG
authors.lastname: Zaphiropoulos
mesh_terms: D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D053823:Hedgehog Proteins / Q000378:metabolism; D006801:Humans; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D015513:Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D012319:RNA Polymerase II / Q000378:metabolism; D022661:RNA, Untranslated / Q000235:genetics*; D015398:Signal Transduction; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation; D016212:Transforming Growth Factor beta / Q000378:metabolism; D000071676:Zinc Finger Protein GLI1
chemical_list: D002843:Chromatin; D053823:Hedgehog Proteins; D015513:Oncogene Proteins; D022661:RNA, Untranslated; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D000071676:Zinc Finger Protein GLI1; D012319:RNA Polymerase II
keywords: Cancer; Chromatin; GLI1; Gene expression; Non-coding RNA; Transcription
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