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Accurate detection of Asparagine Synthetase (ASNS) using quantitative real-time PCR (qRT-PCR), without requiring DNaseI treatment

Leukemia volume 19, pages 2368–2370 (2005)Cite this article

TO THE EDITOR

Asparagine auxotrophy, which forms the basis for treatment of leukemia with L-asparaginase, (L-asp) has been shown in animal tumor cells to be related to the loss of expression of the enzyme Asparagine Synthetase (ASNS) (EC 6.3.5.4).1 The extension of this concept to L-asp sensitivity in clinical cases of acute lymphoblastic leukaemia (ALL) has recently been challenged by the results of ASNS mRNA measurements in patients with t(12;21) translocations. The methodology favoured for measuring ASNS expression has been quantitative real-time PCR (qRT-PCR).2, 3, 4 This technique presents numerous difficulties in application, particularly in the normalization of results and the use of reference genes, as recently reviewed.5 Indeed, using variations of the method, a number of workers have drawn differing conclusions from studies of leukemic cells in short-term culture and in marrow specimens.2, 3, 4 In addition, an examination of the mRNA and genomic sequences for ASNS shows additional factors that could contribute to the uncertainty. We therefore propose a simplified method for this assay in which these features are taken into account.

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References

  1. 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. Br J Cancer 2001; 85: 930–935.

    Article  CAS  Google Scholar 

  2. 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 

  3. Irino T, Kitoh T, Koami K, Kashima T, Mukai K, Takeuchi E et al. Establishment of real-time polymerase chain reaction method for quantitative analysis of asparagine synthetase expression. J Mol Diagn 2004; 6: 217–224.

    Article  CAS  Google Scholar 

  4. 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 

  5. Bustin SA . Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol 2002; 29: 23–39.

    Article  CAS  Google Scholar 

  6. de Kok JB, Roelofs RW, Giesendorf BA, Pennings JL, Waas ET, Feuth T et al. Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes. Lab Invest 2005; 85: 154–159.

    Article  CAS  Google Scholar 

  7. Hurteau GJ, Spivack SD . mRNA-specific reverse transcription-polymerase chain reaction from human tissue extracts. Anal Biochem 2002; 307: 304–315.

    Article  CAS  Google Scholar 

  8. 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 

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  1. Ordway Research Institute, Albany, NY, USA

    G J Hurteau, J D Broome & G J Brock

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  1. G J Hurteau
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  2. J D Broome
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Correspondence to G J Brock.

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Hurteau, G., Broome, J. & Brock, G. Accurate detection of Asparagine Synthetase (ASNS) using quantitative real-time PCR (qRT-PCR), without requiring DNaseI treatment. Leukemia 19, 2368–2370 (2005). https://doi.org/10.1038/sj.leu.2403969

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