Skip to main content
SpringerLink
Log in
Book cover

Hematologic Malignancies pp 83–116Cite as

Molecular Genetic Methods in Diagnosis and Treatment

  • Protocol

  • 400 Accesses

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 55))

Abstract

During the past 20 yr, molecular biology and molecular genetics have provided techniques and procedures that allow the precise diagnoses of leukemias and lymphomas (for a review, see ref. 1). These methodologies have provided powerful, robust, and precise characterizations of genetic mutations and gene rearrangements (both normal and abnormal) that have been applied to the diagnosis and treatment of these malignancies. In this chapter we discuss methodologies currently utilized within the molecular diagnostic laboratory of cancer centers and major academic hospitals. We introduce these methodologies in both general terms and within the context of a differential diagnosis of leukemia/lymphoma. The long-term goal of the molecular hematologist/oncologist is to describe all mutations that occur within a given type or subtype of leukemia/lymphoma. This involves the discovery of new somatic and possibly hereditary mutations that contribute to the genesis of these hematologic malignancies. However, for this information to be useful in treatment regimens, therapies that target or affect these growth-dysregulating mutations must be found. Because diagnosis of mutation and gene rearrangement in the hematologic malignancies is far ahead of therapeutic modalities, we also discuss new research tools that will have impact on the experimental hematologist.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Kinniburgh, A. J. and Ward, P. M. (1977) The application of molecular genetics and biology to the diagnosis and treatment of cancer. Adv. Oncol. 13, 3–8.

    Google Scholar 

  2. Tonegawa, S. (1983) Somatic generation of antibody diversity. Nature 302, 575–581.

    Article  CAS  PubMed  Google Scholar 

  3. Hood, L., Kronenberg, M., and Hunkapiller, T. (1985) T cell antigen receptors and the immunoglobulin supergene family. Cell 40, 225–229.

    Article  CAS  PubMed  Google Scholar 

  4. Korsmeyer, S. J. (1988) B-lymphoid neoplasias: immunoglobulin genes are molecular determinants of clonality lineage, differentiation and translocation. Adv. Intern. Med. 33, 1–15.

    CAS  PubMed  Google Scholar 

  5. van Dongen, J. M. J. and Wolvers-Tettero, I. L. M. (1991) Analysis of immunoglobulin and T cell receptor genes. Part 1: basic and technical aspects. Clin. Chim. Acta 198, 1–92.

    Article  PubMed  Google Scholar 

  6. Hieter, P. A., Korsmeyer, S. J., Waldmann, T. A., et al. (1981) Human immunoglobulin K light-chain genes are deleted or rearranged in lambda-producing B cells. Nature 290, 369–372.

    Article  Google Scholar 

  7. Groffen, J., Stephenson, J. R., Hersterkamp, N., et al. (1990) Philadelphia chromosome breakpoints are clustered within a limited region, bcr, on chromosome 22. Cell 36, 93–99.

    Article  Google Scholar 

  8. Sawyers, C. L., Timson, L., Kawasaki, E. S., et al. (1990) Molecular relapse in chronic myelogenous leukemia patients after bone marrow transplantation detect by polymerase chain reaction. Proc. Natl. Acad. Sci. USA 87, 563–567.

    Article  CAS  PubMed  Google Scholar 

  9. Yunis, J. J. (1983) The chromosomal basis of human neoplasia. Science 221, 227–236.

    Article  CAS  PubMed  Google Scholar 

  10. Levine, E. G., Arthur, D. C., Fizzera, G., et al. (1985) There are differences in cytogenetic abnormalities among histologic subtypes of non-Hodgkin’s lympoma. Blood 66, 1414–1422.

    CAS  PubMed  Google Scholar 

  11. Lipford, E., Wright, J. J., Urba, W., et al. (1987) Refinement of lymphoma cytogenetics by the chromosome 18q21 major breakpoint region. Blood 70, 1816–1823.

    CAS  PubMed  Google Scholar 

  12. Bakhshi, A., Wright, J. J., Graininger, W., et al. (1987) Mechanism of the t(14;18) chromosomal translocation: structural analyses of both derivative 14 and 18 reciprocal partners. Proc. Natl. Acad. Sci. USA 84, 2396–2400.

    Article  CAS  PubMed  Google Scholar 

  13. Lee, M. S., Chang, K. S., Cabanillas, F., et al. (1987) Detection of minimal residual cells carrying the t(14;18) by DNA sequence amplification. Science 237, 175–178.

    Article  CAS  PubMed  Google Scholar 

  14. Ngan, B. Y., Nourse, J., and Cleary, M. L. (1989) Detection of chromosomal translocation t(14; 18) within the minor cluster region of bcl-2 by polymerase chain reaction and direct genomic sequencing of the enzymatically amplified DNA in follicular lymphomas. Blood 73, 1759–1762.

    CAS  PubMed  Google Scholar 

  15. Gribben, J. G., Freedman, A. S., Woo, S. D., et al. (1991) All advanced stage nonHodgkin’s lymphomas with a polymerase chain reaction amplifiable breakpoint of bcl-2 have residual cells containing the bcl-2 rearrangement at evaluation and after treatment. Blood 78, 3275–3280.

    CAS  PubMed  Google Scholar 

  16. Gribben, J. G., Neuberg, D., Barber, M., et al. (1994) Detection of residual lymphoma cells by polymerase chain reaction in peripheral blood is significantly less predictive for relapse than detection in bone marrow. Blood 83, 3800–3807.

    CAS  PubMed  Google Scholar 

  17. Pezzella, F. Ralfkiaer, E., Gatter, K. C., and Mason, D. Y. (1990) The 14;18 translocation in European cases of follicular lymphoma: comparison of southern blotting and the polymerase chain reaction. Br. J. Haematol. 76, 58–64.

    Article  CAS  PubMed  Google Scholar 

  18. Tsujimoto, Y., Bashir, M. M., Givol, I., et al. (1987) DNA rearrangements in human follicular lymphoma can involve the 5? or 3? region of the bcl-2 gene. Proc. Natl. Acad. Sci. USA 84, 1329–1331.

    Article  CAS  PubMed  Google Scholar 

  19. Pezzella, F., Jones, M., Ralfkiaer, E., et al. (1992) Evaluation of bcl-2 protein expression and 14; 18 translocation as prognostic markers in follicular lymphoma. Br. J. Cancer 65, 87–89.

    Article  CAS  PubMed  Google Scholar 

  20. Gribben, J. G., Neuberg, D., Freedman, A. S., et al. (1993) Detection by poly-merase chain reaction of residual cells with bcl-2 translocation is associated with increased risk of relapse after autologous bone marrowtransplantation for B-cell lymphoma. Blood 81, 3449–3457.

    CAS  PubMed  Google Scholar 

  21. Moore, J. S., Friedman, D. F., Silberstein, L. E., et al. (1995) Clinical heterogene-ity reflects biologic diversity in chronic lymphocytic leukemia. Crit. Rev. Oncol. Hematol. 20, 141–164.

    Article  CAS  PubMed  Google Scholar 

  22. Schena, M., Gottardi, D., Ghia, P., et al. (1993) The role of Bcl-2 in the pathogenesis of Bcl-2 in the pathogenesis of B chronic lymphocytic leukemia. Leuk. Lymphoma 11, 173–179.

    Article  CAS  PubMed  Google Scholar 

  23. Hanada, M., Delia, D., Aiello, A., et al. (1993) Bcl-2 gene hypomethylation and highlevel expression in B-cell chronic lymphocytic leukemia. Blood 82, 1820–1828.

    CAS  PubMed  Google Scholar 

  24. Sawyers, C. L., Timson, L., Kawaski, E. S., et al. (1990) Molecular relapse in chronic myelogenous leukemia patients after bone marrow transplantation detected by polymerase chain reaction. Proc. Natl. Acad. Sci. USA 87, 563–567.

    Article  CAS  PubMed  Google Scholar 

  25. Mitani, K. (1996) Chromosomal abnormalities and oncogenes. Int. J. Hematol. 63, 81–93.

    Article  CAS  PubMed  Google Scholar 

  26. Drexler, H. G., Borkhardt, A., and Jansen, J. W. (1995) Detection of chromosomal translocation in leukemia-lymphoma cells by polymerase chain reaction. Leuk. Lymphoma 19, 59–80.

    Article  Google Scholar 

  27. Miyake, K., Inokuchi, K., Dan, K., and Nomura, T. (1993) Alterations in the deleted in colorectal cancer gene in human, primary leukemia. Blood 82, 927–930.

    CAS  PubMed  Google Scholar 

  28. Velculescu, V. E., Zhang, L., Vogelstein, B., and Kinzler, K. W. (1995) Serial analysis of gene expression. Science 270, 484–487.

    Article  CAS  PubMed  Google Scholar 

  29. Zhang, L., Zhow, W., Velculescu, V. E., Hern, S. E., Hruban, R. H., Hamilton, S. R., Vogelstein, B., and Kinzler, K. W. (1997) Gene expression profiles in normal and cancer cells. Science 276, 1268–1272.

    Article  CAS  PubMed  Google Scholar 

  30. Del Sal, G., Manfiolette, G., and Schneider, C. (1989) The CTAB-DNA precipitation method: a common mini-scale preparation of template DNA from phagemids, phages or plasmids suitable for sequencing. Biotechniques 7, 514–519.

    PubMed  Google Scholar 

  31. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  32. Beishuizen, A. Verhoeven, M.-A. J., Mol, E. J., Breit, T. M., WolversTettero, I. L. M., and van Dongen, J. J. M. (1993) Detection of immunoglobulin heavy-chain gene rearrangements by southern blot analysis: recommendations for optimal results. Leukemia 7, 2045–2053.

    CAS  PubMed  Google Scholar 

  33. Silva, A. J., Johnson, J. P., and White, R. L. (1987) Characterization of a highly polymorphic region 5? to JH in the human immunoglobulin heavy chain. Nucleic Acid Res. 15, 2845–2857.

    Article  Google Scholar 

  34. Limpens, J., de Jong, D., van Krieken, J. H., et al. (1991) Bc1-2/JH rearrangements in benign lymphoid tissues with follicular hyperplasia. Oncogene 6, 2271–2276.

    CAS  PubMed  Google Scholar 

  35. Limpens, J., Stad, R., Vos, C., et al. (1995) Lymphoma-associated translocation t(14;18) in blood B cells of normal individuals. Blood 85, 2528–2536.

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Molecular Diagnostics, Roswell Park Cancer Institute, Buffalo, NY

    Pamela Ward

  2. Department of Human Genetics, Roswell Park Cancer Institute, Buffalo, NY

    Alan Kinniburgh

Authors
  1. Pamela Ward
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alan Kinniburgh
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. VA Medical Center and the Medical College of Georgia, Augusta, GA

    Guy B. Faguet

Rights and permissions

Reprints and permissions

Copyright information

© 2001 Humana Press Inc.

About this protocol

Cite this protocol

Ward, P., Kinniburgh, A. (2001). Molecular Genetic Methods in Diagnosis and Treatment. In: Faguet, G.B. (eds) Hematologic Malignancies. Methods in Molecular Medicine™, vol 55. Humana Press. https://doi.org/10.1385/1-59259-074-8:83

Download citation

  • DOI: https://doi.org/10.1385/1-59259-074-8:83

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-543-0

  • Online ISBN: 978-1-59259-074-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation