title: A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT.
abstract: BACKGROUND
  We attempted to clone candidate genes on 10p 14-15 which may regulate hTERT expression, through exon trapping using 3 BAC clones covering the region. After obtaining 20 exons, we examined the function of RGM249 (RGM: RNA gene for miRNAs) we cloned from primary cultured human hepatocytes and hepatoma cell lines. We confirmed approximately 20 bp products digested by Dicer, and investigated the function of this cloned gene and its involvement in hTERT expression by transfecting the hepatoma cell lines with full-length dsRNA, gene-specific designed siRNA, and shRNA-generating plasmid.
  
  
  RESULTS
  RGM249 showed cancer-dominant intense expression similar to hTERT in cancer cell lines, whereas very weak expression was evident in human primary hepatocytes without telomerase activity. This gene was predicted to be a noncoding precursor RNA gene. Interestingly, RGM249 dsRNA, siRNA, and shRNA inhibited more than 80% of hTERT mRNA expression. In contrast, primary cultured cells overexpressing the gene showed no significant change in hTERT mRNA expression; the overexpression of the gene strongly suppressed hTERT mRNA in poorly differentiated cells.
  
  
  CONCLUSIONS
  These findings indicate that RGM249 might be a microRNA precursor gene involved in the differentiation and function upstream of hTERT.
authors.affiliation: Division of Pharmacotherapeutics, Department of Pathophysiological and Therapeutic Science, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8503, Japan. mnmiura@hotmail.com
authors.forename: Norimasa
authors.initials: N
authors.lastname: Miura
authors.forename: Reina
authors.initials: R
authors.lastname: Sato
authors.forename: Tomoe
authors.initials: T
authors.lastname: Tsukamoto
authors.forename: Mika
authors.initials: M
authors.lastname: Shimizu
authors.forename: Hiroko
authors.initials: H
authors.lastname: Kabashima
authors.forename: Miho
authors.initials: M
authors.lastname: Takeda
authors.forename: Shunsaku
authors.initials: S
authors.lastname: Takahashi
authors.forename: Tomomi
authors.initials: T
authors.lastname: Harada
authors.forename: James E
authors.initials: JE
authors.lastname: West
authors.forename: Harry
authors.initials: H
authors.lastname: Drabkin
authors.forename: Jose E
authors.initials: JE
authors.lastname: Mejia
authors.forename: Goshi
authors.initials: G
authors.lastname: Shiota
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Murawaki
authors.forename: Arvind
authors.initials: A
authors.lastname: Virmani
authors.forename: Adi F
authors.initials: AF
authors.lastname: Gazdar
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Oshimura
authors.forename: Junichi
authors.initials: J
authors.lastname: Hasegawa
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D045744:Cell Line, Tumor; D002879:Chromosomes, Human, Pair 10; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019098:Telomerase