A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT

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...

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Bibliographic Details
Published in:BMC molecular biology 2009-02, Vol.10 (1), p.5-5
Main Authors: Miura, Norimasa, Sato, Reina, Tsukamoto, Tomoe, Shimizu, Mika, Kabashima, Hiroko, Takeda, Miho, Takahashi, Shunsaku, Harada, Tomomi, West, James E, Drabkin, Harry, Mejia, Jose E, Shiota, Goshi, Murawaki, Yoshikazu, Virmani, Arvind, Gazdar, Adi F, Oshimura, Mitsuo, Hasegawa, Junichi
Format: Article
Language:English
Subjects:
Base Sequence
Cell Line
Cell Line, Tumor
Chromosomes, Human, Pair 10
Gene expression
Genetic aspects
Humans
MicroRNAs - genetics
MicroRNAs - metabolism
Molecular Sequence Data
Oligonucleotide Array Sequence Analysis
Physiological aspects
Reverse transcriptase
Ribosomal RNA
RNA Interference
RNA, Messenger - metabolism
RNA, Small Interfering - genetics
RNA, Small Interfering - metabolism
Telomerase - genetics
Telomerase - metabolism
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Summary: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. 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. These findings indicate that RGM249 might be a microRNA precursor gene involved in the differentiation and function upstream of hTERT.
ISSN:1471-2199
1471-2199
DOI:10.1186/1471-2199-10-5