Multiplex PCR for Identifying BCR-ABL Fusion Transcript Types in Northeastern Thailand Chronic Myeloid Leukemia Patients

Authors

  • Kanokon Chootawiriyasakul Department of Medicine, Faculty of Medicine, Khon Kaen University
  • Kanchana Chansung Department of Medicine, Faculty of Medicine, Khon Kaen University

Keywords:

Multiplex PCR; BCR-ABL Fusion Transcript Types; Chronic Myeloid Leukemia (CML)

Abstract

Background and Objective:  Philadelphia chromosome is the chromosome originating from the reciprocal translocation between long arms of chromosome 9 and chromosome 22 t (9;22) (q34; q11), it presents in 90 - 95% of patients with chronic myeloid leukemia. The aberration results from a reciprocal translocation are creating a BCR-ABL fusion gene. There are two major forms of the BCR-ABL fusion gene, involving ABL exon 2, but including different exons of BCR gene. The transcript b3a2 and b2a2 codes for a p210 protein. Other fusion gene leads to the expression of an e1a2 transcript, which codes for a p190 protein. Its frequency varies in different populations but there are no data from northeastern Thailand. In this study we aimed to report BCR-ABL fusion transcript types in northeastern Thailand CML patients by Multiple Reverse Transcriptase-Polymerase Chain Reaction (Multiplex PCR).

Materials and Methods Retrospective descriptive and analytical study of all adult CML patients in Srinagarind and Khon Kaen hospitals, which is the main tertiary medical center in the northeastern region Thailand. Data has been collected from medical records for 3 years (January 2017 to January 2020).   Peripheral blood or bone marrow samples were analyzed by multiplex PCR from 177 adult northeastern Thailand CML patients. Aim to identify BCR-ABL fusion transcript types in northeastern Thailand CML patients by Multiplex PCR.

Results: All patients examined were positive for some type of BCR/ABL rearrangement. The majority of the patients (93.79%) expressed one of the p210 BCR-ABL transcripts, b3a2 and b2a2 transcript types were detected in 58.19% and 35.59% respectively. Co-expression of b3a2 (p210)/e1a2 (p230) was detected in 0.56%. The expression of an e1a2 transcript type, which codes for a p190 protein showed 4.52%. And 1.14% was detected in e19a2 transcript type. Co-expression of p210/p190 was not detected.

Conclusions: Multiplex RT-PCR is useful and saves time in the detection of BCR-ABL fusion transcript types; the occurrence of these transcripts associated with CML can assist in prognosis and treatment of disease.

References

1. Rowley JD. A new consistent chromosomal abnormality in chronic myelogenous leukemia identified by quinacrine fluorescence and Giemsa staining. Nature 1973; 243: 290-293.
2. Léglise MCH, Pluchon-Rivière E, Geneviève Le Calvez JF, Berthou CH, Autrand C, Luc Sensebè DB, et al. Molecular diagnosis and follow-up in myeloproliferative syndromes and acute leukemias: Correlation between expression of fusion transcripts and disease progression in chronic myeloid leukemia. Leuk Lymphoma 1996; 21: 187-199.
3. Crist W, Carrol A, Shuster J, Jackson J, Head D, Borowitz M, et al. Philadelphia chromosome-positive childhood acute lymphoblastic leukemia: Clinical and cytogenetic characteristics and treatment outcome: A Pediatric Oncology Group (POG) study. Blood 1990; 76: 489-494.
4. Melo JV. The diversity of BCR-ABL fusion proteins and their relationship to leukemia phenotype. Blood 1996; 88: 2375-2384.
5. Shtivelman E, Lifshitz B, Gale RP, Canaani E. Fused transcript of abl and bcr genes in chronic myelogenous leukaemia. Nature 1985; 315:550-554.
6. Ben-Neriah Y, Daley GQ, Mes-Masson AM, Witte ON Baltimore D. The chronic myelogenous leukemiaspecific p210 protein is the product of the bcrabl hybrid gene. Science 1986; 233:212-214.
7. Okamoto K, Karasawa M, Sakai H, Ogura H, Morita K, Naruse T. A novel acute lymphoid leukemia type BCRABL transcript in chronic myelogenous leukaemia. Br J Haematol 1997; 96:611-613.
8. Lichty BD, Keating A, Callum J, Yee K, Croxford R, Corpus G, et al. Expression of p210 and p190 BCR-ABL due to alternative splicing in chronic myelogenous leukaemia. Br J Haematol 1998; 103:711-415.
9. Saglio G, Pane F, Gottardi E, Frigeri MR. Consistent amounts of acute leukemia-associated P190 BCR/ABL transcripts are expressed by chronic myelogenous leukemia patients at diagnosis. Blood 1996; 87:1075-1080.
10. Pane F, Frigeri F, Sindona M, Luciano L, Ferrara F, Cimino R, et al. Neutrophilic chronic myeloid leukemia: A distinct disease with a specific molecular marker (BCR-ABL with C3/A2 junction). Blood 1996; 88:2410-2414.
11. MittreH, Leymaire P, Macro M, Leporrier M. A new case of chronic myeloid leukemia with c3-a2 BCR-ABL junction. Is it really a distinct disease. Blood 1997; 89:4239-4241.
12. Haskovec C, Pozetto C, Polak J, Maritano D, Serra A, Klamova H, et al. P230 BCR-ABL protein may be associated with an acute leukemia phenotype. Br J Haematol 1998; 103:1104-1108.
13. Lee JJ, Kim HJ, Kim YJ, Lee S, Hwang JY, Kim YL, et al. Imatinib induces a cytogenetic response in blast crisis or interferon failure chronic myeloid leukemia patients with e19a2 BCR-ABL transcripts. Leukemia 2004; 18:1539-1540.
14. T. R. Randolph. Myeloproliferative neoplasms. In: E. M. Keohane, L. J. Smith, J.M. Walenga, editors. Clinical Principles and Applications. 5th ed. Saunders: Rodak’s Hematology; 2016: 561–590.
15. Goh HG, Hwang JY, Kim SH, Lee YH, Kim YL, Kim DW. Comprehensive analysis of BCR-ABL transcript types in Korean CML patients using a newly developed multiplex RT-PCR. Transl Res 2006; 148: 249-256.
16. Mondal BC, Bandyopadhyay A, Majumdar S, Mukhopadhyay A, Chandra S, Chaudhuri U, et al. Molecular profiling of chronic myeloid leukemia in eastern India. Am J Hematol 2006; 81: 845-849.
17. Balatzenko G, Vundinti BR, Margarita G. Correlation between the type of bcr-abl transcripts and blood cell counts in chronic myeloid leukemia – a possible influence of mdr1 gene expression. HR [Internet]. 23Mar.2011 [cited 20Feb.2020]; 3(1): e3. Available from: https://www.pagepress.org/journals/index.php/hr/article/view/hr.2011.e3
18. Kawasaki ES, Clark SS, Coyne MY, Smith SD, Champlin R, Witte ON, Mc Cormick FP. Diagnosis of chronic myeloid and acute lymphocytic leukemia by detection of leukemia-specific mRNA sequences amplified in vitro. Proc Natl Acad Sci USA 1988; 85: 5698-5702.
19. Goh HG, Lin M, Fukushima T, Saglio G, Kim D, Choi SY, et al. Sensitive quantitation of minimal residual disease in chronic myeloid leukemia using nanofluidic digital polymerase chain reaction assay. Leuk Lymphoma 2011; 52: 896-904.
20. Marjan Yaghmaie, Seyed H. Ghaffari, Ardashir Ghavamzadeh, Kamran Alimoghaddam, Mohammad Jahani, Seyed-Asadollah Mousavi, et al., Frequency of BCR-ABL fusion transcripts in Iranian patients with chronic myeloid leukemia. Arch Iran Med. 2008; 11: 247–251.
21. Goh HG, Hwang JY, Kim SH, Lee YH, Kim YL, Kim DW. Comprehensive analysis of BCR-ABL transcript types in Korean CML patients using a newly developed multiplex RT-PCR. Transl Res 2006; 148: 249-256.
22. Mondal BC, Bandyopadhyay A, Majumdar S, Mukhopadhyay A, Chandra S, Chaudhuri U, et al. Molecular profiling of chronic myeloid leukemia in eastern India. Am J Hematol 2006; 81: 845-849.
23. RM Arana‐Trejo E, Ruíz Sánchez G, Ignacio‐Ibarra E, Báez De La Fuente O, Garces E, GÓMEZ Morales, et al. BCR/ABL p210, p190 and p230 fusion genes in 250 Mexican patients with chronic myeloid leukaemia (CML). Clin Lab Haematol 2002; 24(3): 145-150. doi: 10.1046/j.1365-2257.2002.00413.x.

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Published

2020-10-16

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1.
Chootawiriyasakul K, Chansung K. Multiplex PCR for Identifying BCR-ABL Fusion Transcript Types in Northeastern Thailand Chronic Myeloid Leukemia Patients. SRIMEDJ [Internet]. 2020 Oct. 16 [cited 2024 Dec. 23];35(6):720-5. Available from: https://li01.tci-thaijo.org/index.php/SRIMEDJ/article/view/247336

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