Silencing effect of cpg island hypermethylation and histone modifications on o6-methylguanine-dna methyltransferase (mgmt) gene expression in human cancer

ABSTRACT _O_6-methylguanine-DNA methyltransferase (_MGMT_) repairs the cytotoxic and mutagenic _O_6-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. Access through your institution Buy or subscribe This

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METHYLTRANSFERASE ACTIVITY IN COLORECTAL CANCER IS DIRECTED TOWARDS H3K36ME3 MARKED CPG ISLANDS Article Open access 29 January 2021 DNMT3B OVEREXPRESSION DOWNREGULATES GENES WITH CPG

ISLANDS, COMMON MOTIFS, AND TRANSCRIPTION FACTOR BINDING SITES THAT INTERACT WITH DNMT3B Article Open access 02 December 2022 THE GENOME-WIDE MUTATIONAL CONSEQUENCES OF DNA HYPOMETHYLATION

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Download references 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. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Division of Molecular Biology and Genetics, Department of Biomolecular Sciences, Saga

Medical School, 5-1-1 Nabeshima, Saga, 849-8501, Japan Tetsuji Nakagawachi, Hidenobu Soejima, Wei Zhao, Ken Higashimoto, Yuji Satoh, Shiroh Matsukura & Tsunehiro Mukai * Department of

Biochemistry II, Graduate School of Medicine, Nagoya University, Japan Takeshi Urano & Koichi Furukawa * Hokkaido Institute of Public Health, Japan Shinichi Kudo * Department of Surgery,

Saga Medical School, Japan Yoshihiko Kitajima & Kohji Miyazaki * Division of Histology, Department of Pathology, International Medical Center of Japan Research Institute, Japan Haruhito

Harada * Membrane Dynamics Project, Synchrotron Radiation Research Network, Harima Institute at Spring-8, Riken, Japan Hideki Matsuzaki * Department of Molecular Biology, Institute of

Gerontology, Nippon Medical School, Japan Mitsuru Emi * Division of Neurofunctional Genomics, Medical Institute of Bioregulation, Kyushu University and Core Research for Evolutional Science

and Technology, Japan Science and Technology Corporation, Yusaku Nakabeppu * Biomolecular Engineering Research Institute (BERI), Japan Mutsuo Sekiguchi Authors * Tetsuji Nakagawachi View

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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 Download citation * Received: 19 May 2003 * Revised: 04 September 2003 * Accepted: 09 September 2003 * Published: 04

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DNA methylation * histone modification * histone methylation * chromatin * gene silencing