Determinación de la frecuencia de los transcritos de fusión de BCR-ABL (e1a2, b2a2 y  b3a2) por RT-PCR en muestras de médula ósea procedentes de pacientes con leucemia  mieloide crónica  (LMC) y leucemia linfoblástica aguda tipo B (LLA-B)

O. Morales; C. Carranza; P. Saravia

doi:10.54495/Rev.Cientifica.v22i1.117

Authors

O. Morales Escuela de Química Biológica, Facultad de Ciencias Químicas y Farmacia, Universidad de San Carlos de Guatemala (USAC)
C. Carranza Instituto de Investigación y Educación para las Enfermedades Genéticas y Metabólicas (INVEGEM)
P. Saravia Escuela de Química Biológica, Facultad de Ciencias Químicas y Farmacia, Universidad de San Carlos de Guatemala (USAC)

DOI:

https://doi.org/10.54495/Rev.Cientifica.v22i1.117

Keywords:

BCR-ABL, RT-PCR, Chronic myeloid leukemia, type B acute lymphoblastic leukemia

Abstract

At the molecular level translocation t(9;22) (q34, q11), known as the Philadelphia chromosome (Ph +), originates chimeric gene BCR-ABL coding for the expression of three fusion transcripts corresponding to b2a2, b3a2 and e1a2 . It has been shown that there is a relationship between the type of transcript pathological and clinical-demographic characteristics of leukemic patients. In Guatemala, there are no data regarding the frequency of BCR-ABL transcripts in the diagnosis of leukemia, so now the genetic characterization of leukemic Guatemalan population with this condition has been rimpossible to make. The main objective of this study
was to determine the frequency of fusion transcripts BCR-ABL by means of the chain reaction technique of
reverse transcription polymerase (RT-PCR) in bone marrow samples from patients with CML and ALL-B to
help the genetic characterization of leukemic Guatemalan population. In the present study we determined that
the frequency of expression of different transcripts chimeric BCR-ABL gene in 35 bone marrow samples from
patients with CML and ALL-B corresponds to a low frequency for e1a2 transcript (11%) with respect to the
frequency of the b2a2 transcript (43%) and b3a2 (43%) and with a frequency of 3% coexpression represented by b2a2/b3a2 transcripts. Identified the predominance could e1a2 transcript in patients with ALL-B and b2a2 and b3a2 transcripts and their coexpression (b2a2/b3a2) in CML patients, however no significant differences in the frequency of expression of b2a2 and b3a2 compared (P= 0.369). It was found that the findings of studies conducted in different geographical regions of the world for patients with ALL-B match with the frequency reported in the Guatemalan in leukemic population with this condition detected in 100% of patients with transcript expression pathological e1a2. We conclude that the frequency of BCR-ABL transcripts between the Guatemalan population CML agrees with that reported in studies conducted in Latin American countries, where the population is mainly mestizo group and is inverse to that reported in Western and Eastern countries where there is predominance of Caucasian and Asian populations, because in such studies frequently reported a significant difference between the expression frequency of b2a2 (30-40%) compared with b3a2 expression (50-60%). This suggests that the Guatemalan population expressing leukemic fusion transcripts of chimeric BCR-ABL gene has a different biological behavior compared to that reported in Asian and Caucasian population product of genetic variability among populations as a possible explanation for the differences observed in this study.

Downloads

Download data is not yet available.

References

Adler, R., Vielhmann, D., Kuhlisch, E., Martiniak, Y., Rottgers, S., Harbott, J. et al. (2009). Correlation of BCR-ABL transcript variants with patients characteristics in childhood chronic myeloid leukemia. European Journal of Haematology, 82, 112-118. https://doi.org/10.1111/j.1600-0609.2008.01170.x DOI: https://doi.org/10.1111/j.1600-0609.2008.01170.x

Almaguer, C. (2003). Interpretación clínica de la biometría hemática. Medicina Universitaria, 5, 35-40.

Arana, R., Sánchez, E., Ignacio, G., De La Fuente, E., Garces, O., Morales, E. et al. (2002). BCR-ABL p210, p190 and p230 fusion genes in 250 Mexican patients with chronic myeloid leukemia (CML). Clinical & Laboratory Haematology, 24, 145–150. https://doi.org/10.1046/j.1365-2257.2002.00413.x DOI: https://doi.org/10.1046/j.1365-2257.2002.00413.x

Aurer, I., Butturini, A., & Gale, R. (1991). BCR-ABL rearrangements in children with Philadelphia chromosome-positive chronic myelogenous leucemia. Blood, 78, 2407-10. https://doi.org/10.1182/blood.V78.9.2407.2407 DOI: https://doi.org/10.1182/blood.V78.9.2407.bloodjournal7892407

Balatzenko, G., Vundinti, B., & Guenova, M. (2001). Correlation between the type of BCR-ABL transcripts and blood cell counts in chronic myeloid leukemia: a possible influence of mdr1 gene expression. Hematology Reports, 3:e3, 5-8. https://doi.org/10.4081/hr.2011.e3 DOI: https://doi.org/10.4081/hr.2011.e3

Benjamin, L., Ebert, A., & Todd, R. (2004). Genomic approaches to hematologic malignancies. Blood, 104, 923-932. https://doi.org/10.1182/blood-2004-01-0274 DOI: https://doi.org/10.1182/blood-2004-01-0274

Bianchi, C., Cairoli, R., Marenco, P., Muti, G., Del Monte, U., & Perego, R. (1995). Detection by polymerase chain reaction of BCR-ABL transcripts in myeloproliferative diseases at time of diagnosis and for monitoring chronic myelogenous leukemia patients after bone marrow transplantation. European Journal of Cancer, 31A, 197– 201. https://doi.org/10.1016/0959-8049(94)00449-F DOI: https://doi.org/10.1016/0959-8049(94)00449-F

Branford, S., Hughes, T., & Rudzki, Z. (2002). Dual transcription of b2a2 and b3a2 BCR-ABL transcripts in chronic myeloid leukaemia is confined to patients with a linked polymorphism within the BCR gene. British Journal of Haematology, 117, 875-877. https://doi.org/10.1046/j.1365-2141.2002.03508.x DOI: https://doi.org/10.1046/j.1365-2141.2002.03508.x

Cerveira, N., Ferreira, S., Dória, S., Veiga, I., Ferreira, F., Mariz, J. et al. (2000). Detection of prognostic significant translocation in childhood acute lymphoblastic leukemia by one-step multiplex reverse transcription polymerase chain reaction, British Journal of Haematology, 109, 638-640. https://doi.org/10.1046/j.1365-2141.2000.02051.x DOI: https://doi.org/10.1046/j.1365-2141.2000.02051.x

Dobrovic, A., Trainor, K., & Morley, A. (1988). Detection of the molecular abnormality in chronic myeloid leukemia by use of the polymerase chain reaction. Blood, 72, 2063-2065. https://doi.org/10.1182/blood.V72.6.2063.2063 DOI: https://doi.org/10.1182/blood.V72.6.2063.2063

de Klein, A., Van Kessel, A., Grosveld, G., Bartram C., Hagemeijer, A., Bootsma, D. et al. (1982). A cellular oncogene is translocated to the Philadelphia chromosome in chronic myelocytic leukemia. Nature, 300, 765-767. https://doi.org/10.1038/300765a0 DOI: https://doi.org/10.1038/300765a0

de Lemos, J., de Oliveira, C., Scerni, A., Bentes, A. et al. (2005). Differential molecular response of the transcripts B2A2 and B3A2 to imatinib mesylate in chronic myeloid leukemia. Genetical Molecular Research, 4, 803-811.

Eisenberg, A., Silver, R., Soper, L., Arlin, Z., Coleman, M., Bernhardt, B. et al. (1988). The location of breakpoints within the breakpoint cluster region (bcr) of chromosome 22 in chronic myeloid leukemia. Leukemia, 2, 642-647.

Emad I., Osman, & Kamal, H. (2010). Frequencies of BCR-ABL fusion transcripts among Sudanese chronic myeloid leukemia patients. Genetics and Molecular Biology, 33, 229-231. https://doi.org/10.1590/S1415-47572010005000037 DOI: https://doi.org/10.1590/S1415-47572010005000037

Fialkow, P., Jacobson, R., & Papayannopoulou, T. (1977). Chronic myelocytic leukemia: clonal origin a stem cell common to the granulocyte, erythrocyte, platelet and monocyte/macrophage. American Journal of Medicine, 63, 125-130. https://doi.org/10.1016/0002-9343(77)90124-3 DOI: https://doi.org/10.1016/0002-9343(77)90124-3

Goh, H., Hwang, J., Kim, S., Lee, Y., Kim, Y, & Kim, D. (2006). Comprehensive analysis of BCR-ABL transcript types in Korean CML patients using a newly developed multiplex RT-PCR. Translational Research, 248, 249-56. https://doi.org/10.1016/j.trsl.2006.07.002 DOI: https://doi.org/10.1016/j.trsl.2006.07.002

Hasan, SK., Sazawal, S., Kumar, B., Chaubey, R., Mishra, P., Mir, R. et al. (2006). Childhood CML in India: b2a2 transcript is more common than b3a2. Cancer genetic Cytogenetic, 169, 76-7. https://doi.org/10.1016/j.cancergencyto.2006.01.015 DOI: https://doi.org/10.1016/j.cancergencyto.2006.01.015

Heisterkamp, N., Stam, K., Groffen, J., de Klein, A., & Grosveld, G. (1985). Structural organization of the BCR gene and its role in the Ph translocation. Nature, 315, 758-761. https://doi.org/10.1038/315758a0 DOI: https://doi.org/10.1038/315758a0

Henegariu, O., Heerema, N., Dlouhy, S., Vance, G., & Vogt, P. (1997). Multiplex PCR: critical parameters and step by step protocol. Biotechniques, 23, 504–511. https://doi.org/10.2144/97233rr01 DOI: https://doi.org/10.2144/97233rr01

Hermans, A. Heisterkamp, N., von Lindern, M., van Baal, S., Meije, D., van der Plas, D. et al. (1987). Unique fusion of BCR and ABL genes in Philadelphia chromosome positive acute lymphoblastic leukemia. Cell, 51, 33-44. https://doi.org/10.1016/0092-8674(87)90007-9 DOI: https://doi.org/10.1016/0092-8674(87)90007-9

Herrera, P., Calbacho, M., Heras, C., Ramos, M., López, J., & Odriozola, J. (2006). Monitorización de la enfermedad mínima residual en la leucemia mieloide crónica mediante la reacción en cadena de la polimerasa en tiempo real. Methods and Findings in Experimental & Clinical Pharmacology, 28, 47-54.

Ito, T., Tanaka, H., Tanaka, K., Ito, K., Kyo, T., Dohy, H. et al. (2004). Insertion of a genomic fragment of chromosome 19 between BCR intron 19 and ABL intron 1a in a chronic myeloid leukaemia patient with BCR-ABL (e19a2) transcript, British Journal of Haematology, 126, 750–5. https://doi.org/10.1111/j.1365-2141.2004.05119.x DOI: https://doi.org/10.1111/j.1365-2141.2004.05119.x

Lee, A., Kirk, J., Edmans, S., & Radich, J. (1995) Multiplex PCR of BCR/ABL fusion transcripts in Philadelphia positive acute lymphoblastic leukemia, Genome Research, 4, 283-287. https://doi.org/10.1101/gr.4.5.283 DOI: https://doi.org/10.1101/gr.4.5.283

Lee, M., Le Maistre, A., Kantarjian, H., Talpaz, M., Freireich, E., Trujillo, J. et al. (1989) Detection of two alternative BCR-ABL mRNA junctions and minimal residual disease in Philadelphia chromosome positive chronic myelogenous leukemia by polymerase chain reaction. Blood, 73, 2165-2170. https://doi.org/10.1182/blood.V73.8.2165.2165 DOI: https://doi.org/10.1182/blood.V73.8.2165.bloodjournal7382165

Lichty, B., Keating, A., Callum, J., Yee,K., Croxford, R.,Corpus, G. et al. (1998). Expression of p210 and p190 BCR-ABL due to alternative splicing in chronic myelogenous leukaemia. British Journal of Haematology, 103, 711-715. https://doi.org/10.1046/j.1365-2141.1998.01033.x DOI: https://doi.org/10.1046/j.1365-2141.1998.01033.x

Martiat, P., Ifrah, N., Rassool, F., Morgan, G. et al. (1989). Molecular analysis of Philadelphia positive essential thrombocythemia. Leukemia, 3,563-565.

Melo, J. (1996). The diversity of BCR-ABL fusion proteins and their relationship to leukemia phenotype. Blood, 88, 2375. https://doi.org/10.1182/blood.V88.7.2375.bloodjournal8872375 DOI: https://doi.org/10.1182/blood.V88.7.2375.bloodjournal8872375

Meza, J., Gutiérrez, M., Vásquez, A., Delgado, J., Esparza, M., Gonzáles, J. et al. (2007) Prevalence of the BCR-ABL1 transcripts in Mexican patients with chronic myelogenus leucemia. Review of Clinical Investigation, 59, 338-341.

Milis, K., Benn, P., & Birnie, G. (1991) Does the breakpoint within the major breakpoint cluster region (M-bcr) influence the duration of the chronic phase in chronic myeloid leukemia? An analytical comparison of current literature. Blood, 78, 1155-1161. https://doi.org/10.1182/blood.V78.5.1155.1155 DOI: https://doi.org/10.1182/blood.V78.5.1155.1155

Millot, F., Traore, P., Guilhot, J., Nelken, B., Leblanc, T., Leverger, G. et al. (2005). Clinical and biological features at diagnosis in 40 children with chronic myeloid leukemia. Pediatrics, 116, 140-3. https://doi.org/10.1542/peds.2004-2473 DOI: https://doi.org/10.1542/peds.2004-2473

Mondal, B., Bandyopadhyay, A., Majumdar, S., Mukhopadhyay, A., Chandra, S., Chaudhuri, U. et al. (2006). Molecular Profiling of Chronic Myeloid Leukemia in Eastern India, American Journal of Hematology, 81, 845–849. https://doi.org/10.1002/ajh.20682 DOI: https://doi.org/10.1002/ajh.20682

Nowell, P., & Hungerford, D. (1960) A minute chromosome in human chronic granulocytic leukemia. Science, 132, 1497.

Olopade, I., & Pichert G. (2001). Cancer genetics in oncology practice. Annals of Oncology, 12, 895-908. https://doi.org/10.1023/A:1011176107455 DOI: https://doi.org/10.1023/A:1011176107455

Opalka, B., Wandl, U., Stutenkemper, R., Kloke, O. et al. (1992). No correlation between the type of Bcr-Abl hybrid messenger RNA and platelet counts in chronic myelogenous leukemia. Blood, 80, 1854-1855. https://doi.org/10.1182/blood.V80.7.1854.1854 DOI: https://doi.org/10.1182/blood.V80.7.1854.1854

Paz, C., Burgo, R., Morillo, S., Santos, J., Fiallo, B., & Leone, P. (2002). BCR-ABL rearrangement frequencies in chronic myeloid leukemia and acute lymphoblastic leukemia in Ecuador, South America. Cancer Genetics & Cytogenetics, 132, 65-67. https://doi.org/10.1016/S0165-4608(01)00515-5 DOI: https://doi.org/10.1016/S0165-4608(01)00515-5

Perego, R., Costantini, M., Cornacchini, G., Gargantini, L., Bianchi, C., Pungolino, E. et al. (2000). The possible influences of b2a2 and b3a2 BCR-ABL protein structure on thrombopoiesis in chronic myeloid leukemia. European Journal of Cancer, 36, 1395–1401. https://doi.org/10.1016/S0959-8049(00)00128-3 DOI: https://doi.org/10.1016/S0959-8049(00)00128-3

Rosas, A., Martinez, M., Ayala, M., Vela, J., Bahena, P., Vadillo, M. et al. (2003). Análisis del tipo de transcrito BCR-ABL y su relación con la cuenta plaquetaria en pacientes mexicanos con leucemia mieloide crónica. Gaceta Médica de México, 139, 553-559.

Rozman, C., Urbano, A., Cervantes, F., Rozman, M., Colomer, D., Féliz, P. et al. (1995). Analysis of the clinical relevance of the breakpoint location within M-bcr and the type of chimeric mRNA in chronic myelogenous leukemia. Leukemia, 9, 1104-1107.

Ruiz, G., Garcés, J., Reyes, V., & Ruiz, G. (2004). Frequencies of the breakpoint cluster region types of the BCR-ABL fusion gene in Mexican Mestizo patients with chronic myelogenous leukemia. Journal of Clinical Research, 56, 605-8

Shepherd, P., Bond, C., & Allan, N. (1992). Molecular breakpoints and platelet counts in chronic myeloid leukemia. Blood, 80, 556-557. https://doi.org/10.1182/blood.V80.2.556.556 DOI: https://doi.org/10.1182/blood.V80.2.556.bloodjournal802556

Shepherd, P., Suffolk, R., Halsey, J., & Allan, N. (1995). Analysis of molecular breakpoint and m-RNA transcript in a prospective randomized trial of interferon in chronic mueloid leukaemia: no correlation with clinical features, cytogenetic response, duration of chronic phase, or survival, British Journal of Haematology, 89, 546-54. https://doi.org/10.1111/j.1365-2141.1995.tb08362.x DOI: https://doi.org/10.1111/j.1365-2141.1995.tb08362.x

Udomsakdi-Auewarakul, C., U-Pratya, Y., Boonmoh, S., & Vatanavicharn, S. (2000). Detection of molecular variants of BCR-ABL gene in bone marrow and blood of patients with chronic myeloid leukemia by reverse-transcriptase polymerase chain reaction (RT-PCR). Journal of the Medicine Association of Thailand, 83, 928 –35.

van Rhee, F., Hochhaus, A., Lin, F., Melo, J., Goldman, J., & Cross, N. (1996) p190 BCR-ABL mRNA is expressed at low levels in p210-positive chronic myeloid and acute lymphoblastic leukemias. Blood, 87, 5213–5217. https://doi.org/10.1182/blood.V87.12.5213.bloodjournal87125213 DOI: https://doi.org/10.1182/blood.V87.12.5213.bloodjournal87125213

Yaghmaie, M., Ghaffari, S., Ghavamzadeh, A., Alimoghaddam, K., Jahani, M., Mousavi, S. et al. (2008). Frequency of BCR-ABL Fusion Transcripts in Iranian Patients with Chronic Myeloid Leukemia. Iranian Archive of Medicine, 11, 247 – 251.