Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Biotechnical Methods Section BTS
  • Published:

Biotechnical Methods Section (BTS)

Detection of methylthioadenosine phosphorylase (MTAP) and p16 gene deletion in T cell acute lymphoblastic leukemia by real-time quantitative PCR assay

Leukemia volume 14, pages 935–940 (2000)Cite this article

Abstract

Methylthioadenosine phosphorylase (MTAP) deficiency in tumors can be therapeutically exploited for selective therapy. Many tumors lacking MTAP have been found to homozygously delete the chromosome 9p region containing the p16 tumor suppressor gene. Several methods have been used to detect chromosome 9p deletions in primary tumors. However, the accurate diagnosis of chromosome 9p deletions has been hampered by the presence of contaminating normal cells. In search of an accurate and sensitive diagnostic method, we have developed the real-time polymerase chain reaction assay using the TaqMan chemistry for quantitative detection of MTAP and p16 gene deletions. The assay's feasibility was tested with peripheral blood leukocytes (PBL) from 29 patients with adult T cell leukemia (ATL) previously analyzed with Southern blot analysis and validated on 39 PBL or bone marrow samples from childhood T cell acute lymphoblastic leukemia (T-ALL). Homozygous deletions of MTAP and p16 genes were detected respectively in six (20.7%) and eight (27.6%) of 29 ATL samples and in 15 (38.5%) and 23 (59%) of 39 T-ALL samples. The results correlated well with those of Southern blot analysis. It is of significance that the newly developed method can successfully detect homozygous deletions of these genes in samples containing as low as 33% blast cells. This rapid and sensitive method may be useful in searching for candidates for selective therapy targeting MTAP deficiency.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  1. Batova A, Diccianni MB, Omura-Minamisawa M, Yu J, Carrera CJ, Bridgeman LJ, Kung FH, Pullen J, Amylon MD, Yu AL . Use of alanosine as a methylthioadenosine phosphorylase-selective therapy for T-cell acute lymphoblastic leukemia in vitro Cancer Res 1999 59: 1492–1497

    CAS  PubMed  Google Scholar 

  2. Kamatani N, Nelson-Rees WA, Carson DA . Selective killing of human malignant cell lines deficient in methylthioadenosine phosphorylase, a purine metabolic enzyme Proc Natl Acad Sci USA 1981 78: 1219–1223

    Article  CAS  Google Scholar 

  3. Batova A, Diccianni MB, Nobori T, Vu T, Yu J, Bridgeman L, Yu AL . Frequent deletion in the methylthioadenosine phosphorylase gene in T-cell acute lymphoblastic leukemia: strategies for enzyme-targeted therapy Blood 1996 88: 3083–3090

    CAS  PubMed  Google Scholar 

  4. Pegg AE, Williams-Ashman HG . Phosphate-stimulated breakdown of 5′-methylthioadenosine by rat ventral prostate Biochem J 1969 115: 241–247

    Article  CAS  PubMed Central  Google Scholar 

  5. Backlund PS Jr, Smith RA . Methionine synthesis from 5′-methylthioadenosine in rat liver J Biol Chem 1981 256: 1533–1535

    PubMed  Google Scholar 

  6. Kamatani N, Carson DA . Dependence of adenine production upon polyamine synthesis in cultured human lymphoblasts Biochim Biophys Acta 1981 675: 344–350

    Article  CAS  Google Scholar 

  7. Toohey JI . Methylthioadenosine nucleoside phosphorylase deficiency in methylthio-dependent cancer cells Biochem Biophys Res Commun 1978 83: 27–35

    Article  CAS  Google Scholar 

  8. Kamatani N, Carson DA . Abnormal regulation of methylthioadenosine and polyamine metabolism in methylthioadenosine phosphorylase-deficient human leukemic cell lines Cancer Res 1980 40: 4178–4182

    CAS  PubMed  Google Scholar 

  9. Fitchen JH, Riscoe MK, Dana BW, Lawrence JH, Ferro AJ . Methylthioadenosine phosphorylase deficiency in human leukemias and solid tumors Cancer Res 1986 46: 5409–5412

    CAS  PubMed  Google Scholar 

  10. Nobori T, Karras JG, Della Ragione F, Waltz TA, Chen PP, Carson DA . Absence of methylthioadenosine phosphorylase in human gliomas Cancer Res 1991 51: 3193–3197

    CAS  PubMed  Google Scholar 

  11. Nobori T, Szinai I, Amox D, Parker B, Olopade OI, Buchhagen DL, Carson DA . Methylthioadenosine phosphorylase deficiency in human non-small cell lung cancers Cancer Res 1993 53: 1098–1101

    CAS  PubMed  Google Scholar 

  12. Sahota A, Webster DR, Potter CF, Simmonds HA, Rodgers AV, Gibson T . Methylthioadenosine phosphorylase activity in human erythrocytes Clin Chim Acta 1983 128: 283–290

    Article  CAS  Google Scholar 

  13. Yu J, Batova A, Shao L, Carrera CJ, Yu AL . Presence of methylthioadenosine phosphorylase (MTAP) in hematopoietic stem/progenitor cells: its therapeutic implication for MTAP(−) malignancies Clin Cancer Res 1997 3: 433–438

    CAS  PubMed  Google Scholar 

  14. Hori H, Tran P, Carrera CJ, Hori Y, Rosenbach MD, Carson DA, Nobori T . Methylthioadenosine phosphorylase cDNA transfection alters sensitivity to depletion of purine and methionine in A549 lung cancer cells Cancer Res 1996 56: 5653–5658

    CAS  PubMed  Google Scholar 

  15. Chen ZH, Zhang H, Savarese TM . Gene deletion chemoselectivity: codeletion of the genes for p16INK4, methylthioadenosine phosphorylase, and the α- and β-interferons in human pancreatic cell carcinoma lines and its implications for chemotherapy Cancer Res 1996 56: 1083–1090

    CAS  PubMed  Google Scholar 

  16. Chen ZH, Olopade OI, Savarese TM . Expression of methylthioadenosine phosphorylase cDNA in p16-, MTAP-malignant cells: restoration of methylthioadenosine phosphorylase-dependent salvage pathways and alterations of sensitivity to inhibitors of purine de novo synthesis Mol Pharmacol 1997 52: 903–911

    Article  CAS  Google Scholar 

  17. Kamb A, Gruis NA, Weaver-Feldhaus J, Liu Q, Harshman K, Tavtigian SV, Stockert E, Day RS III, Johnson BE, Skolnick MH . A cell cycle regulator potentially involved in genesis of many tumor types Science 1994 264: 436–440

    Article  CAS  Google Scholar 

  18. Nobori T, Miura K, Wu DJ, Lois A, Takabayashi K, Carson DA . Deletions of the cyclin-dependent kinase-4 inhibitor gene in multiple human cancers Nature 1994 368: 753–756

    Article  CAS  Google Scholar 

  19. Nobori T, Takabayashi K, Tran P, Orvis L, Batova A, Yu AL, Carson DA . Genomic cloning of methylthioadenosine phosphorylase: a purine metabolic enzyme deficient in multiple different cancers Proc Natl Acad Sci USA 1996 93: 6203–6208

    Article  CAS  Google Scholar 

  20. Ogawa S, Hirano N, Sato N, Takahashi T, Hangaishi A, Tanaka K, Kurokawa M, Tanaka T, Mitani K, Yazaki Y, Hirai H . Homozygous loss of the cyclin-dependent kinase 4-inhibitor (p16) gene in human leukemias Blood 1994 84: 2431–2435

    CAS  PubMed  Google Scholar 

  21. Walker DG, Duan W, Popovic EA, Kaye AH, Tomlinson FH, Lavin M . Homozygous deletions of the multiple tumor suppressor gene 1 in the progression of human astrocytomas Cancer Res 1995 55: 20–23

    CAS  PubMed  Google Scholar 

  22. Okamoto A, Hussain SP, Hagiwara K, Spillare EA, Rusin MR, Demetrick DJ, Serrano M, Hannon GJ, Shiseki M, Zariwala M, Xiong Y, Beach DH, Yokota J, Harris CC . Mutations in the p16INK4/MTS1/CDKN2, and p15INK4B/MTS2, and p18 genes in primary and metastatic lung cancer Cancer Res 1995 55: 1448–1451

    CAS  PubMed  Google Scholar 

  23. Okuda T, Shurtleff SA, Valentine MB, Raimondi SC, Head DR, Behm F, Curcio-Brint AM, Liu Q, Pui CH, Sherr CJ, Beach D, Look AT, Downing JR . Frequent deletion of p16INK4a/MTS1 and p15INK4b/MTS2 in pediatric acute lymphoblastic leukemia Blood 1995 85: 2321–2330

    CAS  Google Scholar 

  24. Hori Y, Hori H, Yamada Y, Carrera CJ, Tomonaga M, Kamihira S, Carson DA, Nobori T . The methylthioadenosine phosphorylase gene is frequently co-deleted with the p16INK4a gene in acute type adult T-cell leukemia Int J Cancer 1998 75: 51–56

    Article  CAS  Google Scholar 

  25. Shimoyama M and members of The Lymphoma Study Group . Diagnostic criteria and classification of clinical subtypes of adult T-cell leukaemia-lymphoma. A report from the Lymphoma Study Group (1984–87) Br J Haematol 1991 79: 428–437

    Article  CAS  Google Scholar 

  26. Yamada Y, Hatta Y, Murata K, Sugawara K, Ikeda S, Mine M, Maeda T, Hirakata Y, Kamihira S, Tsukasaki K, Ogawa S, Hirai H, Koeffler HP, Tomonaga M . Deletions of p15 and/or p16 genes as a poor-prognosis factor in adult T-cell leukemia J Clin Oncol 1997 15: 1778–1785

    Article  CAS  Google Scholar 

  27. Hatta Y, Hirama T, Miller CW, Yamada Y, Tomonaga M, Koeffler HP . Homozygous deletions of the p15 (MTS2) and p16 (CDKN2/MTS1) genes in adult T-cell leukemia Blood 1995 85: 2699–2704

    CAS  PubMed  Google Scholar 

  28. Diccianni MB, Batova A, Yu J, Vu T, Pullen J, Amylon M, Pollock BH, Yu AL . Shortened survival after relapse in T-cell acute lymphoblastic leukemia patients with p16/p15 deletions Leukemia Res 1997 21: 549–558

    Article  CAS  Google Scholar 

  29. Perry A, Nobori T, Ru N, Anderl K, Borell TJ, Mohapatra G, Feuerstein BG, Jenkins RB, Carson DA . Detection of p16 gene deletions in gliomas: a comparison of fluorescence in situ hybridization (FISH) versus quantitative PCR J Neuropath Exp Neur 1997 56: 999–1008

    Article  CAS  Google Scholar 

  30. Heid CA, Stevens J, Livak KJ, Williams PM . Real time quantitative PCR Genome Res 1996 6: 986–994

    Article  CAS  Google Scholar 

  31. Lockey C, Otto E, Long Z . Real-time fluorescence detection of a single DNA molecule Biotechniques 1998 24: 744–746

    Article  CAS  Google Scholar 

  32. Tran PT, Hori H, Hori Y, Okumura K, Kagotani K, Taguchi H, Carson DA, Nobori T . Molecular cloning of the human methylthioadenosine phosphorylase processed pseudogene and localization to 3q28 Gene 1997 186: 263–269

    Article  Google Scholar 

  33. Cairns P, Sidransky D . Molecular methods for the diagnosis of cancer Biochim Biophys Acta 1999 1423: C11–C18

    CAS  PubMed  Google Scholar 

  34. Takeuchi S, Koike M, Seriu T, Bartram CR, Slater J, Park S, Miyoshi I, Koeffler HP . Homozygous deletions at 9p21 in childhood acute lymphoblastic leukemia detected by microsatellite analysis Leukemia 1997 11: 1636–1640

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported in part by Grants-in-aid from the Ministry of Education, Science, Sports and Culture of Japan and from the Mie Medical Research Foundation and by a grant from the Okasan-Kato Foundation.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Mie University Hospital, Tsu, Mie, Japan

    TJ M'soka, H Kawasaki & Y Komada

  2. Central Clinical Laboratories, Mie University Hospital, Tsu, Mie, Japan

    J Nishioka, A Taga & K Kato

  3. Department of Laboratory Medicine, Nagasaki University School of Medicine, Nagasaki, Japan

    Y Yamada

  4. Department of Pediatrics, University of California at San Diego, California, USA

    A Yu

  5. Department of Laboratory Medicine, Mie University Hospital, Tsu, Mie, Japan

    T Nobori

Authors
  1. TJ M'soka
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J Nishioka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A Taga
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H Kawasaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Y Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. A Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Y Komada
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. T Nobori
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

M'soka, T., Nishioka, J., Taga, A. et al. Detection of methylthioadenosine phosphorylase (MTAP) and p16 gene deletion in T cell acute lymphoblastic leukemia by real-time quantitative PCR assay. Leukemia 14, 935–940 (2000). https://doi.org/10.1038/sj.leu.2401771

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2401771

Keywords

This article is cited by

Search

Advanced search

Quick links