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

Norimasa Miura, Reina Sato, Tomoe Tsukamoto, Mika Shimizu, Hiroko Kabashima, Miho Takeda, Shunsaku Takahashi, Tomomi Harada, James E. West, Harry Drabkin, Jose E. Mejia, Goshi Shiota, Yoshikazu Murawaki, Arvind Virmani, Adi F. Gazdar, Mitsuo Oshimura, Junichi Hasegawa

Research output: Contribution to journalArticle

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Abstract

Background: We attempted to clone candidate genes on 10p14-15 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. Conclusion: These findings indicate that RGM249 might be a microRNA precursor gene involved in the differentiation and function upstream of hTERT.

Original languageEnglish (US)
Article number5
JournalBMC Molecular Biology
Volume10
DOIs
StatePublished - Feb 2 2009

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Untranslated RNA
Chromosomes
Genes
Small Interfering RNA
MicroRNAs
Cell Line
Messenger RNA
Hepatocytes
Hepatocellular Carcinoma
Exons
Clone Cells
Telomerase
Cultured Cells
Neoplasms
Plasmids
RNA

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Miura, N., Sato, R., Tsukamoto, T., Shimizu, M., Kabashima, H., Takeda, M., ... Hasegawa, J. (2009). A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT. BMC Molecular Biology, 10, [5]. https://doi.org/10.1186/1471-2199-10-5

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

In: BMC Molecular Biology, Vol. 10, 5, 02.02.2009.

Research output: Contribution to journalArticle

Miura, N, Sato, R, Tsukamoto, T, Shimizu, M, Kabashima, H, Takeda, M, Takahashi, S, Harada, T, West, JE, Drabkin, H, Mejia, JE, Shiota, G, Murawaki, Y, Virmani, A, Gazdar, AF, Oshimura, M & Hasegawa, J 2009, 'A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT', BMC Molecular Biology, vol. 10, 5. https://doi.org/10.1186/1471-2199-10-5
Miura N, Sato R, Tsukamoto T, Shimizu M, Kabashima H, Takeda M et al. A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT. BMC Molecular Biology. 2009 Feb 2;10. 5. https://doi.org/10.1186/1471-2199-10-5
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. / A noncoding RNA gene on chromosome 10p15.3 may function upstream of hTERT. In: BMC Molecular Biology. 2009 ; Vol. 10.
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abstract = "Background: We attempted to clone candidate genes on 10p14-15 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. Conclusion: These findings indicate that RGM249 might be a microRNA precursor gene involved in the differentiation and function upstream of hTERT.",
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AU - Kabashima, Hiroko

AU - Takeda, Miho

AU - Takahashi, Shunsaku

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AU - Mejia, Jose E.

AU - Shiota, Goshi

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AU - Virmani, Arvind

AU - Gazdar, Adi F.

AU - Oshimura, Mitsuo

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