Methylation analysis of asparagine synthetase gene in acute lymphoblastic leukemia cells

Access through your institution Buy or subscribe Asparagine synthetase (ASY) is an enzyme that produces asparagine from aspartic acid in an ATP-dependent manner. The _ASY_ gene is conserved

in mammalian cells; however, the expression of the gene is repressed in acute lymphoblastic leukemia (ALL) cells preventing them from synthesizing asparagine.1 As ALL cells depend on

extracellular sources of asparagine to survive, depletion of asparagine by L-asparaginase leads to cell death; therefore, the enzyme has been used for the treatment of childhood ALL.2

Methylation of CpG islands of genes is one of the epigenetic mechanisms for silencing of genes, and aberrant methylation of CpG islands has been observed in a cohort of genes in tumor cells.

Little is known about the methylation status of the _ASY_ gene in ALL patients; here, we show that this gene is often methylated in ALL bone marrow samples. We also examined the methylation

status of _ASY_ gene in brain and breast tumors. Using five brain tumor samples, we analyzed the methylation status in the CpG island of ASY from −313 to +25 by sequencing. Most CpG sites

in the gene were not methylated in brain tumors (Figure 2b). In breast tumors, 11 samples were analyzed by _Bst_UI digestion, and none were digested by _Bst_UI (data not shown), suggesting

that the gene is not methylated in breast tumors. Taken together, these results clearly show that the CpG island of the _ASY_ gene is methylated in both B-lineage ALL and T-ALL, but not in

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local taxes which are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support REFERENCES * Broome

JD . Studies on the mechanism of tumor inhibition by L-asparaginase. Effects of the enzyme on asparagine levels in the blood, normal tissues, and 6C3HED lymphomas of mice: differences in

asparagine formation and utilization in asparaginase-sensitive and -resistant lymphoma cells. _J Exp Med_ 1968; 127: 1055–1072. Article  CAS  Google Scholar  * Pinheiro JP, Boos J . The best

way to use asparaginase in childhood acute lymphatic leukaemia-still to be defined? _Br J Haematol_ 2004; 125: 117–127. Article  Google Scholar  * Peng H, Shen N, Qian L, Sun XL, Koduru P,

Goodwin LO _et al_. Hypermethylation of CpG islands in the mouse asparagine synthetase gene: relationship to asparaginase sensitivity in lymphoma cells. Partial methylation in normal cells.

_Br J Cancer_ 2001; 85: 930–935. Article  CAS  Google Scholar  * Ren Y, Roy S, Ding Y, Iqbal J, Broome JD . Methylation of the asparagine synthetase promoter in human leukemic cell lines is

associated with a specific methyl binding protein. _Oncogene_ 2004; 23: 3953–3961. Article  CAS  Google Scholar  * Ding Y, Li Z, Broome JD . Epigenetic changes in the repression and

induction of asparagine synthetase in human leukemic cell lines. _Leukemia_ 2005; 19: 420–426. Article  CAS  Google Scholar  * Stams WA, den Boer ML, Beverloo HB, Meijerink JP, Stigter RL,

van Wering ER _et al_. Sensitivity to L-asparaginase is not associated with expression levels of asparagine synthetase in t(12;21)+ pediatric ALL. _Blood_ 2003; 101: 2743–2747. Article  CAS

  Google Scholar  * Stams WA, den Boer ML, Holleman A, Appel IM, Beverloo HB, van Wering ER _et al_. Asparagine synthetase expression is linked with L-asparaginase resistance in

TEL-AML1-negative but not TEL-AML1-positive pediatric acute lymphoblastic leukemia. _Blood_ 2005; 105: 4223–4225. Article  CAS  Google Scholar  * Krejci O, Starkova J, Otova B, Madzo J,

Kalinova M, Hrusak O _et al_. Upregulation of asparagine synthetase fails to avert cell cycle arrest induced by L-asparaginase in TEL/AML1-positive leukaemic cells. _Leukemia_ 2004; 18:

434–441. Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by NIH grants as well as Parker Hughes Fund. HPK is the holder of the Mark Goodson endowed

Chair in Oncology Research and is a member of the Jonsson Cancer and Molecular Biology Institute, UCLA. The work is dedicated to the memory of David Golde, a friend and mentor. AUTHOR

INFORMATION Author notes * T Akagi and D Yin: These authors contributed equally to this work AUTHORS AND AFFILIATIONS * Division of Hematology and Oncology, Cedars-Sinai Medical Center, UCLA

School of Medicine, Los Angeles, CA, USA T Akagi, D Yin, N Kawamata, I Wolf, C W Miller & H P Koeffler * Institute of Human Genetics, University of Heidelberg, Heidelberg, Germany C R

Bartram * Department of Hematology and Oncology and Transfusion Medicine, University Hospital ‘Benjamin Franklin’, Berlin, Germany W-K Hofmann Authors * T Akagi View author publications You

can also search for this author inPubMed Google Scholar * D Yin View author publications You can also search for this author inPubMed Google Scholar * N Kawamata View author publications You

can also search for this author inPubMed Google Scholar * C R Bartram View author publications You can also search for this author inPubMed Google Scholar * W-K Hofmann View author

publications You can also search for this author inPubMed Google Scholar * I Wolf View author publications You can also search for this author inPubMed Google Scholar * C W Miller View

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CORRESPONDING AUTHOR Correspondence to T Akagi. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Akagi, T., Yin, D., Kawamata, N. _et al._ Methylation

analysis of asparagine synthetase gene in acute lymphoblastic leukemia cells. _Leukemia_ 20, 1303–1306 (2006). https://doi.org/10.1038/sj.leu.2404216 Download citation * Published: 06 April

2006 * Issue Date: 01 July 2006 * DOI: https://doi.org/10.1038/sj.leu.2404216 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable

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