Abstract
The tumor suppressor Chk2 kinase plays crucial roles in regulating cell-cycle checkpoints and apoptosis following DNA damage. We investigated the expression levels of the genes encoding Chk2 and several cell-cycle regulators in nine cell lines from lymphoid malignancies, including three Hodgkin's lymphoma (HL) lines. We found that all HL cell lines exhibited a drastic reduction in Chk2 expression without any apparent mutation of the Chk2 gene. However, expression of Chk2 in HL cells was restored following treatment with the histone deacetylase inhibitors trichostatin A (TsA) and sodium butyrate (SB), or with the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (5Aza-dC). Chromatin-immunoprecipitation (Chip) assays revealed that treatment of HL cells with TsA, SB or 5Aza-dC resulted in increased levels of acetylated histones H3 and H4, and decreased levels of dimethylated H3 lysine 9 at the Chk2 promoter. These results indicate that expression of the Chk2 gene is downregulated in HL cells via epigenetic mechanisms.
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Abbreviations
- ATM:
-
ataxia-telangiectasia-mutated
- 5Az-dC:
-
5-aza-2′-deoxycytidine
- Chip:
-
chromatin-immunoprecipitation
- CKI:
-
cyclin-dependent protein kinase inhibitor
- FCS:
-
fetal calf serum
- HDAC:
-
histone deacetylase
- HL:
-
Hodgkin's lymphoma
- H/RS:
-
Hodgkin and Reed-Sternberg
- IR:
-
ionizing radiation
- LFS:
-
Li-Fraumeni syndrome
- MSP:
-
methylation-specific PCR
- PMSF:
-
phenylmethyl sulfonyl fluoride
- SB:
-
sodium butyrate
- TsA:
-
trichostatin A
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Acknowledgements
We thank Drs. M Lamphier and A Noda for critical reading of the manuscript, E Tatsumi for providing HDLM2, KM-H2, L428, KM3, RL, FL318, FL518 and Black93 cells, A Kosugi for providing Jurkat cells and M Takao for encouraging this study. This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science and Technology of Japan, the Yasuda Medical Research Foundation, the Cell Science Research Foundation, Nippon Boehringer Ingelheim, Co., Ltd, and Daiichi Pharmaceutical Co., Ltd.
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Kato, N., Fujimoto, H., Yoda, A. et al. Regulation of Chk2 gene expression in lymphoid malignancies: involvement of epigenetic mechanisms in Hodgkin's lymphoma cell lines. Cell Death Differ 11 (Suppl 2), S153–S161 (2004). https://doi.org/10.1038/sj.cdd.4401461
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DOI: https://doi.org/10.1038/sj.cdd.4401461
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