Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Identification of an epigenetically silenced gene, RFX1, in human glioma cells using restriction landmark genomic scanning

Abstract

To identify the CpG islands differentially methylated in human glioma, we performed restriction landmark genomic scanning with a CpG methylation-sensitive enzyme. We found 12 spots, the intensity of which was entirely lost or decreased in both the human glioma tissues examined as compared with that in matched normal lymphocytes, indicating aberrant methylation of these CpG islands in gliomas. The expression of RFX1, one of the genes associated with the methylated CpG islands, was frequently decreased in human glioma cell lines and tissues. We also demonstrated that the isolated CpG island located in the seventh intron of the RFX1 gene had enhancer activity and was hypermethylated in all of the glioma tissues and cell lines analysed, but not in normal brains or lymphocytes. Treatment of glioma cells with a demethylating agent, 5-azacytidine, resulted in the expression of RFX1, indicating that the silencing of the RFX1 gene may be attributable to its methylation. RFX1 has been implicated in transcriptional downregulation of the proto-oncogene c-myc. By expression of the RFX1 gene, the cellular proliferative activity of glioma cells was suppressed. Taken together, these results suggest that the RFX1 gene may be epigenetically silenced in human gliomas and involved in glioma tumorigenesis.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

References

  • Blin N and Stafford DW . (1976). Nucleic Acids Res., 3, 2303–2308.

  • Chen L, Smith L, Johnson MR, Wang K, Diasio RB and Smith JB . (2000). J. Biol. Chem., 275, 32227–32233.

  • Costello JF, Fruhwald MC, Smiraglia DJ, Rush LJ, Robertson GP, Gao X, Wright FA, Feramisco JD, Peltomaki P, Lang JC, Schuller DE, Yu L, Bloomfield CD, Caligiuri MA, Yates A, Nishikawa R, Su Huang H, Petrelli NJ, Zhang X, O’Dorisio MS, Held WA, Cavenee WK and Plass C . (2000). Nat. Genet., 24, 132–138.

  • Di Cristofano A, Pesce B, Cordon-Cardo C and Pandolfi PP . (1998). Nat. Genet., 19, 348–355.

  • Emery P, Durand B, Mach B and Reith W . (1996). Nucleic Acids Res., 24, 803–807.

  • Frommer M, McDonald LE, Millar DS, Collis CM, Watt F, Grigg GW, Molloy PL and Paul CL . (1992). Proc. Natl. Acad. Sci. USA, 89, 1827–1831.

  • Fults D, Brockmeyer D, Tullous MW, Pedone CA and Cawthon RM . (1992). Cancer Res., 52, 674–679.

  • Gajiwala KS, Chen H, Cornille F, Roques BP, Reith W, Mach B and Burley SK . (2000). Nature, 403, 916–921.

  • Gardiner-Garden M and Frommer M . (1987). J. Mol. Biol., 196, 261–282.

  • Hatada I, Hayashizaki Y, Hirotsune S, Komatsubara H and Mukai T . (1991). Proc. Natl. Acad. Sci. USA, 88, 9523–9527.

  • Humphrey PA, Wong AJ, Vogelstein B, Friedman HS, Werner MH, Bigner DD and Bigner SH . (1988). Cancer Res., 48, 2231–2238.

  • Itano O, Ueda M, Kikuchi K, Shimazu M, Kitagawa Y, Aiura K and Kitajima M . (2000). Oncogene, 19, 1676–1683.

  • Katan-Khaykovich Y and Shaul Y . (2001). Eur. J. Biochem., 268, 3108–3116.

  • Lindsay S and Bird AP . (1987). Nature, 327, 336–338.

  • Maegawa S, Yoshioka H, Itaba N, Kubota N, Nishihara S, Shirayoshi Y, Nanba E and Oshimura M . (2001). Mol. Carcinogen., 31, 1–9.

  • Meehan RR, Lewis JD and Bird AP . (1992). Nucleic Acids Res., 20, 5085–5092.

  • Melki JR, Vincent PC and Clark SJ . (1999). Cancer Res., 59, 3730–3740.

  • Nakamura M, Yonekawa Y, Kleihues P and Ohgaki H . (2001). Lab. Invest., 81, 77–82.

  • Rasheed BK, Wiltshire RN, Bigner SH and Bigner DD . (1999). Curr. Opin. Oncol., 11, 162–167.

  • Saito Y, Kanai Y, Sakamoto M, Saito H, Ishii H and Hirohashi S . (2001). Hepatology, 33, 561–568.

  • Sekita N, Suzuki H, Ichikawa T, Kito H, Akakura K, Igarashi T, Nakayama T, Watanabe M, Shiraishi T, Toyota M, Yoshie O and Ito H . (2001). Jpn. J. Cancer Res., 92, 947–951.

  • Sidransky D, Mikkelsen T, Schwechheimer K, Rosenblum ML, Cavanee W and Vogelstein B . (1992). Nature, 355, 846–847.

  • Siegrist CA, Durand B, Emery P, David E, Hearing P, Mach B and Reith W . (1993). Mol. Cell. Biol., 13, 6375–6384.

  • Smiraglia DJ, Fruhwald MC, Costello JF, McCormick SP, Dai Z, Peltomaki P, O’Dorisio MS, Cavenee WK and Plass C . (1999). Genomics, 58, 254–262.

  • Umezawa A, Yamamoto H, Rhodes K, Klemsz MJ, Maki RA and Oshima RG . (1997). Mol. Cell. Biol., 17, 4885–4894.

  • Whang YE, Wu X, Suzuki H, Reiter RE, Tran C, Vessella RL, Said JW, Isaacs WB and Sawyers CL . (1998). Proc. Natl. Acad. Sci. USA, 95, 5246–5250.

  • Yamada KM and Araki M . (2001). J. Cell Sci., 114, 2375–2382.

  • Zardo G, Tiirikainen MI, Hong C, Misra A, Feuerstein BG, Volik S, Collins CC, Lamborn KR, Bollen A, Pinkel D, Albertson DG and Costello JF . (2002). Nat. Genet., 32, 453–458.

Download references

Acknowledgements

We thank Mr Muramatsu and Mr Iizuka (Keio University School of Medicine) for their technical support. We are also grateful to Dr W Reith (University of Geneva Medical School) for providing us with the pHRFX1 plasmid. This work was supported by grants from grant-in-aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology (MT), the Keio University Medical Science Fund (MT), and grand-in-aid for Scientific Research on Priority Areas from the Japanese Ministry of Education, Culture, Sports, Science and Technology (MU).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Masahiro Toda.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ohashi, Y., Ueda, M., Kawase, T. et al. Identification of an epigenetically silenced gene, RFX1, in human glioma cells using restriction landmark genomic scanning. Oncogene 23, 7772–7779 (2004). https://doi.org/10.1038/sj.onc.1208058

Download citation

  • Received:

  • Revised:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1208058

Keywords

  • glioma
  • RLGS
  • RFX1
  • methylation
  • epigenetics

Further reading

Search

Quick links