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  • Original Paper
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Silencing effect of CpG island hypermethylation and histone modifications on O6-methylguanine-DNA methyltransferase (MGMT) gene expression in human cancer

Abstract

O6-methylguanine-DNA methyltransferase (MGMT) repairs the cytotoxic and mutagenic O6-alkylguanine produced by alkylating agents such as chemotherapeutic agents and mutagens. Recent studies have shown that in a subset of tumors, MGMT expression is inversely linked to hypermethylation of the CpG island in the promoter region; however, how the epigenetic silencing mechanism works, as it relates to hypermethylation, was still unclear. To understand the mechanism, we examined the detailed methylation status of the whole island with bisulfite-sequencing in 19 MGMT non-expressed cancer cell lines. We found two highly methylated regions in the island. One was upstream of exon 1, including minimal promoter, and the other was downstream, including enhancer. Reporter gene assay showed that methylation of both the upstream and downstream regions suppressed luciferase activity drastically. Chromatin immunoprecipitation assay revealed that histone H3 lysine 9 was hypermethylated throughout the island in the MGMT negative line, whereas acetylation on H3 and H4 and methylation on H3 lysine 4 were at significantly high levels outside the minimal promoter in the MGMT-expressed line. Furthermore, MeCP2 preferentially bound to the CpG-methylated island in the MGMT negative line. Given these results, we propose a model for gene silencing of MGMT that is dependent on the epigenetic state in cancer.

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References

  • Bachman KE, Park BH, Rhee I, Rajagopalan H, Herman JG, Baylin SB, Kinzler KW and Vogelstein B . (2003). Cancer Cell, 3, 89–95.

  • Ballestar E and Wolffe AP . (2001). Eur. J. Biochem., 268, 1–6.

  • Baylin SB and Herman JG . (2000). Trends Genet., 16, 168–174.

  • Becker K, Dosch J, Gregel CM, Martin BA and Kaina B . (1996). Cancer Res., 56, 3244–3249.

  • Boggs BA, Cheung P, Heard E, Spector DL, Chinault AC and Allis CD . (2002). Nat. Genet., 30, 73–76.

  • Chen FY, Harris LC, Remack JS and Brent TP . (1997). Proc. Natl. Acad. Sci., 94, 4348–4353.

  • Costello JF, Futscher BW, Tano K, Graunke DM and Pieper RO . (1994a). J. Biol. Chem., 269, 17228–17237.

  • Costello JF, Futscher BW, Kroes RA and Pieper RO . (1994b). Mol. Cell. Biol., 14, 6515–6521.

  • Danam RP, Howell SR, Remack JS and Brent TP . (2001). Int. J. Oncol., 18, 1187–1193.

  • Danam RP, Qian XC, Howell SR and Brent TP . (1999). Mol. Carcinogen., 24, 85–89.

  • Esteller M, Hamilton SR, Burger PC, Baylin SB and Herman JG . (1999). Cancer Res., 59, 793–797.

  • Esteller M, Risques RA, Toyota M, Capella G, Moreno V, Peinado MA, Baylin SB and Herman JG . (2001). Cancer Res., 61, 4689–4692.

  • Esteller M, Toyota M, Sanchez-Cespedes M, Capella G, Peinado MA, Watkins DN, Issa JP, Sidransky D, Baylin SB and Herman JG . (2000). Cancer Res., 60, 2368–2371.

  • Fahrner JA, Eguchi S, Herman JG and Baylin SB . (2002). Cancer Res., 62, 7213–7218.

  • Fuks F, Hurd PJ, Wolf D, Nan X, Bird AP and Kouzarides T . (2003). J. Biol. Chem., 278, 4035–4040.

  • Hall IM, Shankaranarayana GD, Noma K, Ayoub N, Cohen A and Grewal SI . (2002). Science, 297, 2232–2237.

  • Harris LC, Potter PM, Tano K, Shiota S, Mitra S and Brent TP . (1991). Nucleic Acids Res., 16, 6163–6167.

  • Harris LC, Remack JS and Brent TP . (1994). Nucleic acids Res., 22, 4614–4619.

  • Heard E, Rougeulle C, Arnaud D, Avner P, Allis CD and Spector DL . (2001). Cell, 107, 727–738.

  • Herfarth KK, Brent TP, Danam RP, Remack JS, Kodner IJ, Wells Jr SA and Goodfellow PJ . (1999). Mol. Carcinogen., 24, 90–98.

  • Hsieh CL . (1997). Mol. Cell. Biol., 17, 5897–5904.

  • Irvine RA, Lin IG and Hsieh CL . (2002). Mol. Cell. Biol., 22, 6689–6696.

  • Jackson JP, Lindroth AM, Cao X and Jacobsen SE . (2002). Nature, 416, 556–560.

  • Jenuwein T and Allis CD . (2001). Science, 293, 1074–1080.

  • Kawate H, Sakumi K, Tsuzuki T, Nakatsuru Y, Ishikawa T, Takahashi S, Takano H, Noda T and Sekiguchi M . (1998). Proc. Natl. Acad. Sci., 95, 5116–5120.

  • Kondo Y, Shen L and Issa JP . (2003). Mol. Cell. Biol., 23, 206–215.

  • Kudo S . (1998). Mol. Cell. Biol., 18, 5492–5499.

  • Magdinier F and Wolffe AP . (2001). Proc. Natl. Acad. Sci. USA, 98, 4990–4995.

  • Matsukura S, Soejima H, Nakagawachi T, Yakushiji H, Ogawa A, Fukuhara M, Miyazaki K, Nakabeppu Y, Sekiguchi M and Mukai T . (2003). Br. J. Cancer, 88, 521–529.

  • Miller SA, Dykes DD and Polesky HF . (1988). Nucleic Acids Res., 16, 1215.

  • Mukai T and Sekiguchi M . (2002). Oncogene, 21, 9033–9042.

  • Nakayama J, Rice JC, Strahl BD, Allis CD and Grewal SI . (2001). Science, 292, 110–113.

  • Ng HH, Zhang Y, Hendrich B, Johnson CA, Turner BM, Erdjument-Bromage H, Tempst P, Reinberg D and Bird A . (1999). Nat. Genet., 23, 58–61.

  • Nguyen CT, Gonzales FA and Jones PA . (2001). Nucleic Acids Res., 29, 4598–4606.

  • Nguyen CT, Weisenberger DJ, Velicescu M, Gonzales FA, Lin JC, Liang G and Jones PA . (2002). Cancer Res., 32, 6456–6461.

  • Olsson M and Lindahl T . (1980). J. Biol. Chem., 255, 10569–10571.

  • Patel SA, Graunke DM and Pieper RO . (1997). Mol. Cell. Biol., 17, 5813–5822.

  • Paulin R, Grigg GW, Davey MW and Piper AA . (1998). Nucleic Acids Res., 26, 5009–5010.

  • Pegg AE, Wiest L, Foote RS, Mitra S and Perry W . (1983). J. Biol. Chem., 258, 2327–2333.

  • Peters AH, Mermoud JE, O'Carroll D, Pagani M, Schweizer D, Brockdorff N and Jenuwein T . (2002). Nat. Genet., 30, 77–80.

  • Pieper RO, Patel S, Ting SA, Futscher BW and Costello JF . (1996). J. Biol. Chem., 271, 13916–13924.

  • Qian XC and Brent TP . (1997). Cancer Res., 57, 3672–3677.

  • Roder K, Latasa MJ and Sul HS . (2002). J. Biol. Chem., 277, 30543–30550.

  • Sakumi K, Shiraishi A, Shimizu S, Tsuzuki T, Ishikawa T and Sekiguchi M . (1997). Cancer Res., 57, 2415–2418.

  • Satoh Y, Nakagawachi T, Nakadate H, Kaneko Y, Masaki Z, Mukai T and Soejima H . (2003). J. Biochem., 133, 303–308.

  • Tachibana M, Sugimoto K, Fukushima T and Shinkai Y . (2001). J. Biol. Chem., 276, 25309–25317.

  • Tamaru H and Selker EU . (2001). Nature, 414, 277–283.

  • Urano T, Furukawa K and Shiku H . (1993). Oncogene, 8, 1371–1376.

  • von Wronski MA, Harris LC, Tano K, Mitra S, Bigner DD and Brent TP . (1992). Oncol. Res., 4, 167–174.

  • Watts GS, Pieper RO, Costello JF, Peng YM, Dalton WS and Futscher BW . (1997). Mol. Cell. Biol., 17, 5612–5619.

  • Whitehall VL, Walsh MD, Young J, Leggett BA and Jass JR . (2001). Cancer Res., 61, 827–830.

  • Xin Z, Allis CD and Wagstaff J . (2001). Am. J. Hum. Genet., 69, 1389–1394.

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Acknowledgements

We thank Drs M Tachibana and Y Shinkai (Kyoto University, Kyoto) for pGEX4T-G9a-C, K Sugimoto (Osaka Prefecture University, Osaka) for pGEX-H3 (1–57), and M Berne (Tufts University, Boston) for peptide synthesis. We also thank Drs T Abe and M Oka (Yamaguchi University School of Medicine, Ube), Dr Y Shimada (Kyoto University, Kyoto) and Cell Recourse Center for Biomedical Research Institute of Development, Aging & Cancer (Tohoku University, Sendai) for providing cancer cell lines. This work was supported by grants-in-aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS), for COE research from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan, and the Uehara Memorial Foundation.

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Correspondence to Hidenobu Soejima.

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Nakagawachi, T., Soejima, H., Urano, T. et al. Silencing effect of CpG island hypermethylation and histone modifications on O6-methylguanine-DNA methyltransferase (MGMT) gene expression in human cancer. Oncogene 22, 8835–8844 (2003). https://doi.org/10.1038/sj.onc.1207183

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