- Email: [email protected]
Fluorescence in situ hybridization analysis of the cryptic t(12;21) (p13;q22) in childhood B-lineage acute lymphoblastic leukemia
Cancer Genetics and Cytogenetics 132 (2002) 61–64
Short communication
Fluorescence in situ hybridization analysis of the cryptic t(12;21) (p13;q22) in childhood B-lineage acute lymphoblastic leukemia Orly Yehuda-Gafnia, Gabriel Cividallib, Ayala Abrahmovc, Michael Weintrobb, Susana Ben Neriaha, Rachel Cohena, Dvorah Abeliovicha,* a
Department of Human Genetics, Hadassah Hebrew University Hospital and Medical School, Jerusalem 91120, Israel b Department of Pediatrics, Hadassah Hebrew University Hospital and Medical School, Jerusalem 91120, Israel c Department of Pediatrics, Shaare Zedek Medical Center, Jerusalem 91120, Israel Received 7 May 2001; accepted 12 June 2001
Abstract
We evaluated retrospectively the cryptic t(12;21)(p13;q22) in 15 children with early B-lineage acute lymphocytic leukemia who had a normal karyotype by using the locus specific probes of TEL and AML1 genes in a dual color fluorescence in situ hybridization (FISH). The FISH analysis revealed six patients with the fusion gene TEL/AML1 on chromosome 21, three of whom possessed a double fusion gene. In addition, the AML1 probe revealed hyperdiploid clones that were not detected in the conventional cytogenetic analysis. A discrepancy between the proportion of cells with the fusion gene in interphase nuclei and metaphases was noted. © 2002 Elsevier Science Inc. All rights reserved.
1. Introduction The cryptic t(12;21)(p13;q22) is recognized as the most frequent translocation in early B-lineage childhood acute lymphocytic leukemia (ALL) [1]. The results in a fusion transcript of the TEL gene on 12p13 and the AML1 gene on 21q22 [2]. The t(12;21)(p13;q22) cannot be detected by conventional cytogenetics, but can be demonstrated by fluorescence in situ hybridization (FISH) using two cosmids from the TEL and AML1 genes, that detect the fusion gene TEL/AML1 on chromosome 21 or by a polymerase chain reaction (PCR)–based technique [3,4]. This translocation is associated with a favorable prognosis [5]. Several studies have shown that among patients with B-lineage childhood ALL, the t(12;21)(p13;q22) is present in 11–30% of the cases [6–13]. In a retrospective study we analyzed for the t(12;21)(p13;q22) in 15 children with B-lineage ALL, six of whom (40%) had the TEL/AML1 fusion gene. 2. Patients Twenty-two patients received a diagnosis of childhood B-lineage ALL with no evidence of hyperdiploidy between 1994 and 2000. We had material from 15 patients for a retrospective FISH analysis. These cases either had a normal
* Corresponding author. Tel.: 972-2 6776016; fax: 972-2 6777499. E-mail address: [email protected] (D. Abeliovich).
karyotype at diagnosis (n10) or karyotype analysis could not be performed because of poor quality metaphases or low mitotic index (n3); one patient had a translocation and one patient had one cell with trisomy 21. Eight patients were females and seven were males; the age range at diagnosis was 1.5 to 12 years. Eleven children had B-lineage ALL and four had biphenotypic ALL. The clinical description of the patients is presented in Table 1. 3. Cytogenetic and fluorescence in situ hybridization analysis Bone marrow aspirate was processed according to a method previously described [14], using the conditioned medium derived from the human bladder carcinoma cell line 5637. G-banding was performed according to a standard technique [15]. In each case, 10–25 metaphases were analyzed. Fluorescence in situ hybridization analysis was performed with the probes, LSI TEL/AML1 ES (Vysis, Downers Grove, IL, USA) using the fluorochromes SpectrumOrange (AML1) and SpectrumGreen (TEL); blue 4,6diamidino-2-phenylindole-2 HCl (DAPI) was used for counterstaining. Fluorescence in situ hybridization was performed according to the manufacturer’s instructions. Fluorescence microscopy was performed with a Leitz Pleomopak microscope and a cooled monochrome charged-coupled device (CCD) camera in MacProbe FISH analysis (PSI Scientific Systems, League City, TX, USA). In each case, one to five metaphases and at least 50 interphase nuclei were analyzed.
0165-4608/02/$ – see front matter © 2002 Elsevier Science Inc. All rights reserved. PII: S0165-4608(01)00 5 3 0 - 1
62
O. Yehuda-Gafni et al. / Cancer Genetics and Cytogenetics 132 (2002) 61–64
Table 1 The clinical description of the patients Patient no.
Sex
Age at diagnosis (years)
Initial WBC count
1
M
3
7100
2
M
11
63000
3
F
12
9000
4
F
4
7000
5
M
1.5
6700
6
F
3.5
3600
7 8
F F
3.5 12
49000 5000
9 10
M M
3 4
45000 35000
11
M
1.5
7600
12
F
2.2
9500
13
M
3.5
3500
14
F
3
12200
15
F
11.5
101000
Immunophenotype Biphenotypic ALL CD13 , CD33 Pre-B ALL CD10 Pre-B ALL CD10 Early pre-B ALL CD10, CD19, CD34 Early pre-B ALL CD34, CD19, CD10, HLADr Pro pre-B ALL CD34, HLADr preB ALL Biphenotypic ALL
B-lineage ALL Early pre-B ALL CD10, CD19, CD34, HLADr Biphenotypic ALL
pre-B ALL CD10, CD19, CD34, HLADr Early pre-B ALL CD10, CD19, CD34, HLADr Early pre-B ALL CD2, CD7, CD10 Biphenotypic ALL CD19, CD34, CD33, HLADr
Disease outcome Full remission Died Remission Remission
Remission Remission Remission Relapse after two BMT (died) Remission Remission
Remission (no response to steroids) Remission
Remission (no response to steroids) Remission Remission after BMT
Abbreviations: ALL: acute lymphocytic leukemia; BMT: bone marrow transplantation; WBC: white blood cell.
4. Results and discussion The cytogenetic and FISH results are presented in Table 2. Six patients had the TEL/AML1 fusion gene, in three of whom (patients 4, 13, and 14) the fusion gene was duplicated (Fig. 1). Patient 4 had one metaphase with an apparent trisomy 21 and two fusion genes; after the FISH analysis the extra chromosome 21 was interpreted as a derivative of the t(12;21)(p13;q22), an observation encountered by others [12,16,17]. All patients with the cryptic t(12;21)(p13;q22) had pre-B ALL. The age at diagnosis of the patients with the fusion gene TEL/AML1 was between 2.5 and 4 years, and all of them attained remission. In two patients, there was an extra signal of the AML1 cosmid probe with no fusion gene. In these patients, the extra signals represented a state of hyperdiploidy that was not detected in the metaphases analyzed by conventional cytogenetics. In patient 1 with the t(12;17)(p13;q12), the TEL signal remained on chromosome 12 indicating that in this
translocation it is not the rearrangement of the TEL gene that plays a role in the malignant transformation. In some of the patients, the proportion of cells with the TEL/ AML1 fusion gene in metaphases was markedly lower than in interphase nuclei. For example, in patient 4, 2 of 25 metaphases had double signals of the fusion gene; in interphase nuclei it was 30 of 50, and with conventional cytogenetic analysis 1 of 25 cells had the der(21)t(12;21)(p13;q22). In patient 6 the TEL/AML1 fusion gene was noted in 4 of 11 metaphases and in 35 of 50 interphase nuclei, and in patient 13, 2 of 22 cells had the TEL/AML1 fusion gene and in the interphase nuclei it was on 91 of 100. This discrepancy is most likely because of the low mitotic index of the leukemic cells under the culture condition, used as part of the cytogenetic analysis. The relatively high incidence (40%) of patients with the t(12;21)(p13;q22) in our study may be the result of the selection method used for the FISH analysis, namely, patients with pre-B ALL that had normal karyotypes at diagnosis. Our study indicates that in childhood B-lineage ALL,
O. Yehuda-Gafni et al. / Cancer Genetics and Cytogenetics 132 (2002) 61–64
63
Table 2 Cytogenetic and fluorescence in situ hybridization results in bone marrow aspirates of the patients at diagnosis and remission Patient no.
Karyotype [no. of cells]
FISH (metaphase) [no. of cells]
FISH (interphase) [no. of cells]
46,XY,t(12;17)(p13;q12) (46%)/46,XY(54%) [24] 46,XY [15] 46,XX [20]
Normal [8]
Normal [50]
Normal [4] 2XTEL, 3XAML1 (22%)/normal (78%) [9]
4
47,XX,21(4%)/46,XX (96%) [25]
5
46,XY [16]
(TEL/AML1)2 (8%)/normal(92%) [25] 2XTEL, 3XAML2 (40%)/normal (60%) [10]
6
46,XX [18]
7
No metaphases
8 9 10 11 12
46,XX [21] 46,XY [10] 46,XY [20] 46,XY [20] No metaphases
Normal [15] Normal [6] Normal [6] Normal [8] TEL/AML1 [12] Remission: normal [25]
13
46,XY [24]
14
No metaphases
(TEL/AML1)2 (9%)/normal (91%) [22] (TEL/AML1)2 [1] Remission: normal [20]
15
46,XX [16]
Normal [50] 2XTEL, 3XAML1 (20%)/normal (80%) [50] Hyperploidy (TEL/AML1)2 60%)/normal (40%) [50] 2XTEL, 3XAML1 (62%)/normal (38%) [50] Hyperploidy TEL/AML1 (70%)/ normal (30%) [50] TEL/AML1 (98%)/ normal (2%) [50] Remission: normal (98%)/ Tel/AML1 (2%) [100] Normal [50] Normal [50] Normal [50] Normal [50] TEL/AML1 (98%)/ normal (2%) [50] Remission: normal [100] (TEL/AML1) 2 (91%)/normal (9%) [100] (TEL/AML1) 2 (94%)/normal (6%) [50] Remission: normal (99%)/TEL/AML1 (1%) [100] Normal [50]
1 2 3
mainly in pre-B ALL with normal karyotypes the cryptic t(12;21)(p13;q22) may account for a high proportion of patients, and therefore FISH analysis is warranted with special attention to interphase nuclei.
References [1] Romana SP, Poirel H, Leconiat M, Flexor MA, Mauchauffe M, Jonveaux P, Macintyre EA, Berger R, Bernard OA. High frequency of t(12;21) in childhood B-lineage acute lymphoblastic leukemia. Blood 1995;86:4263–9. [2] Golub TR, Barker GF, Bohlander SK, Hiebert SW, Ward DC, BrayWard P, Morgan E, Raimondi SC, Rowley JD, Gilliland GD. Fusion of the TEL gene on 12p13 to the AML1 gene on 21q22 in acute lymphoblastic leukemia. Proc Natl Acad Sci USA 1995;92:4917–21. [3] Romana SP, Le Coniat M, Berger R. t(12;21): a new recurrent translocation in acute lymphoblastic leukemia. Genes Chromosomes Cancer 1994;9:186–91.
TEL/AML1 (36%)/ normal (64%) [11] TEL/AML1 [2] Remission: normal [15]
Normal [16]
[4] Amor DJ, Algar EM, Slater HR, Smith PJ. High frequency of t(12;21) in childhood acute lymphoblastic leukemia detected by RT-PCR. Pathology 1998;30:381–5. [5] Rubnitz JE, Downing JR, Pui CH, Shurtleff SA, Raimondi SC, Evans WE, Head DR, Crist WM, Rivera GK, Hancock ML, Boyett JM, Buijs A, Grosveld G, Behm FG. TEL gene rearrangement in acute lymphoblastic leukemia: a new genetic marker with prognostic significance. J Clin Oncol 1997;15:1150–7. [6] Shurtleff SA, Buijs A, Behm FG, Rubnitz JE, Raimondi SC, Hancock ML, Chan GC, Pui CH, Grosveld G, Downing JR. TEL/AML1 fusion resulting from a cryptic t(12;21) is the most common genetic lesion in pediatric ALL and defines a subgroup of patients with an excellent prognosis. Leukemia 1995;9:1985–9. [7] Liang DC, Chou TB, Chen JS, Shurtleff SA, Rubnitz JE, Downing JR, Pui CH, Shih LY. High incidence of TEL/AML1 fusion resulting from a cryptic t(12;21) in childhood B-lineage acute lymphoblastic leukemia in Taiwan. Leukemia 1996;10:991–3. [8] McLean TW, Ringold S, Neuberg D, Stegmaier K, Tantravahi R, Ritz J, Koeffler HP, Takeuchi S, Janssen JW, Seriu T, Bartram CR, Sallan SE, Gilliland DG, Golub TR. TEL/AML-1 dimerizes and is associ-
64
O. Yehuda-Gafni et al. / Cancer Genetics and Cytogenetics 132 (2002) 61–64
[9]
[10]
[11]
[12]
[13]
[14]
[15]
[16] [17]
Fig. 1. Fluorescence in situ hybridization analysis of patient 13 using TEL (green) and AML1 (red) cosmid probes. (a) Cells with no t(12;21)(p13; q22). (b) Cells with t(12;21)(p13;q22); two der(21)t(12;21)(p13;q22) chromosomes (arrowheads). The arrows points to the residual signal of the AML1 cosmid probe on the derivative chromosome 12.
ated with a favorable outcome in childhood acute lymphoblastic leukemia. Blood 1996;88:4252–8. Harbott J, Viehmann S, Borkhardt A, Henze G, Lampert F. Incidence of TEL/AML1 fusion gene analyzed consecutively in children with acute lymphoblastic leukemia in relapse. Blood 1997;90:4933–7. Eguchi-Ishimae M, Eguchi M, Tanaka K, Hamamoto K, Ohki M, Ueda K, Kamada N. Fluorescence in situ hybridization analysis of 12; 21 translocation in Japanese childhood acute lymphoblastic leukemia. Jpn J Cancer Res 1998;89:783–8. Kempski H, Chalker J, Chessells J, Sturt N, Brickell P, Webb J, Clink JM, Reeves B. An investigation of the t(12;21) rearrangement in children with B-precursor acute lymphoblastic leukaemia using cytogenetic and molecular methods. Br J Haematol 1999;105:684–9. Andreasson P, Hoglund M, Bekassay AN, Garwicz S, Heldrup J, Mitelman F, Johansson B. Cytogenetic and FISH studies of a single center consecutive series of 152 childhood acute lymphoblastic leukemias. Eur J Haematol 2000;65:40–51. Jamil A, Theil KS, Kahwash S, Ruymann FB, Klopfenstein KJ. TEL/ AML-1 fusion gene: Its frequency and prognostic significance in childhood acute lymphoblastic leukemia. Cancer Genet Cytogenet 2000;122:73–8. Michaeli J, Lerer I, Rachmilewitz EA, Fibach E. Stimulation of proliferation of human myeloid cells in culture: application for cytogenetic analysis. Blood 1986;68:790–3. Benn PA, Perle MA. Chromosome staining and banding techniques. In: Rooney DE, Czepulkowski BH, editors. Human cytogenetics, constitutional analysis. A practical approach. Vol. 1. Oxford, Washington DC: IRL Press, 1992, pp. 91–118. Berger R. Acute lymphoblastic leukemia and chromosome 21. Cancer Genet Cytogenet 1997;94:8–12. Loncarevic IF, Roitzheim B, Ritterbach J, Viehmann S, Borkhardt A, Lampert F, Harbott J. Trisomy 21 is a recurrent secondary aberration in childhood acute lymphoblastic leukemia with TEL/AML1 gene fusion. Genes Chromosomes Cancer 1999;24:272–7.
Short communication
Fluorescence in situ hybridization analysis of the cryptic t(12;21) (p13;q22) in childhood B-lineage acute lymphoblastic leukemia Orly Yehuda-Gafnia, Gabriel Cividallib, Ayala Abrahmovc, Michael Weintrobb, Susana Ben Neriaha, Rachel Cohena, Dvorah Abeliovicha,* a
Department of Human Genetics, Hadassah Hebrew University Hospital and Medical School, Jerusalem 91120, Israel b Department of Pediatrics, Hadassah Hebrew University Hospital and Medical School, Jerusalem 91120, Israel c Department of Pediatrics, Shaare Zedek Medical Center, Jerusalem 91120, Israel Received 7 May 2001; accepted 12 June 2001
Abstract
We evaluated retrospectively the cryptic t(12;21)(p13;q22) in 15 children with early B-lineage acute lymphocytic leukemia who had a normal karyotype by using the locus specific probes of TEL and AML1 genes in a dual color fluorescence in situ hybridization (FISH). The FISH analysis revealed six patients with the fusion gene TEL/AML1 on chromosome 21, three of whom possessed a double fusion gene. In addition, the AML1 probe revealed hyperdiploid clones that were not detected in the conventional cytogenetic analysis. A discrepancy between the proportion of cells with the fusion gene in interphase nuclei and metaphases was noted. © 2002 Elsevier Science Inc. All rights reserved.
1. Introduction The cryptic t(12;21)(p13;q22) is recognized as the most frequent translocation in early B-lineage childhood acute lymphocytic leukemia (ALL) [1]. The results in a fusion transcript of the TEL gene on 12p13 and the AML1 gene on 21q22 [2]. The t(12;21)(p13;q22) cannot be detected by conventional cytogenetics, but can be demonstrated by fluorescence in situ hybridization (FISH) using two cosmids from the TEL and AML1 genes, that detect the fusion gene TEL/AML1 on chromosome 21 or by a polymerase chain reaction (PCR)–based technique [3,4]. This translocation is associated with a favorable prognosis [5]. Several studies have shown that among patients with B-lineage childhood ALL, the t(12;21)(p13;q22) is present in 11–30% of the cases [6–13]. In a retrospective study we analyzed for the t(12;21)(p13;q22) in 15 children with B-lineage ALL, six of whom (40%) had the TEL/AML1 fusion gene. 2. Patients Twenty-two patients received a diagnosis of childhood B-lineage ALL with no evidence of hyperdiploidy between 1994 and 2000. We had material from 15 patients for a retrospective FISH analysis. These cases either had a normal
* Corresponding author. Tel.: 972-2 6776016; fax: 972-2 6777499. E-mail address: [email protected] (D. Abeliovich).
karyotype at diagnosis (n10) or karyotype analysis could not be performed because of poor quality metaphases or low mitotic index (n3); one patient had a translocation and one patient had one cell with trisomy 21. Eight patients were females and seven were males; the age range at diagnosis was 1.5 to 12 years. Eleven children had B-lineage ALL and four had biphenotypic ALL. The clinical description of the patients is presented in Table 1. 3. Cytogenetic and fluorescence in situ hybridization analysis Bone marrow aspirate was processed according to a method previously described [14], using the conditioned medium derived from the human bladder carcinoma cell line 5637. G-banding was performed according to a standard technique [15]. In each case, 10–25 metaphases were analyzed. Fluorescence in situ hybridization analysis was performed with the probes, LSI TEL/AML1 ES (Vysis, Downers Grove, IL, USA) using the fluorochromes SpectrumOrange (AML1) and SpectrumGreen (TEL); blue 4,6diamidino-2-phenylindole-2 HCl (DAPI) was used for counterstaining. Fluorescence in situ hybridization was performed according to the manufacturer’s instructions. Fluorescence microscopy was performed with a Leitz Pleomopak microscope and a cooled monochrome charged-coupled device (CCD) camera in MacProbe FISH analysis (PSI Scientific Systems, League City, TX, USA). In each case, one to five metaphases and at least 50 interphase nuclei were analyzed.
0165-4608/02/$ – see front matter © 2002 Elsevier Science Inc. All rights reserved. PII: S0165-4608(01)00 5 3 0 - 1
62
O. Yehuda-Gafni et al. / Cancer Genetics and Cytogenetics 132 (2002) 61–64
Table 1 The clinical description of the patients Patient no.
Sex
Age at diagnosis (years)
Initial WBC count
1
M
3
7100
2
M
11
63000
3
F
12
9000
4
F
4
7000
5
M
1.5
6700
6
F
3.5
3600
7 8
F F
3.5 12
49000 5000
9 10
M M
3 4
45000 35000
11
M
1.5
7600
12
F
2.2
9500
13
M
3.5
3500
14
F
3
12200
15
F
11.5
101000
Immunophenotype Biphenotypic ALL CD13 , CD33 Pre-B ALL CD10 Pre-B ALL CD10 Early pre-B ALL CD10, CD19, CD34 Early pre-B ALL CD34, CD19, CD10, HLADr Pro pre-B ALL CD34, HLADr preB ALL Biphenotypic ALL
B-lineage ALL Early pre-B ALL CD10, CD19, CD34, HLADr Biphenotypic ALL
pre-B ALL CD10, CD19, CD34, HLADr Early pre-B ALL CD10, CD19, CD34, HLADr Early pre-B ALL CD2, CD7, CD10 Biphenotypic ALL CD19, CD34, CD33, HLADr
Disease outcome Full remission Died Remission Remission
Remission Remission Remission Relapse after two BMT (died) Remission Remission
Remission (no response to steroids) Remission
Remission (no response to steroids) Remission Remission after BMT
Abbreviations: ALL: acute lymphocytic leukemia; BMT: bone marrow transplantation; WBC: white blood cell.
4. Results and discussion The cytogenetic and FISH results are presented in Table 2. Six patients had the TEL/AML1 fusion gene, in three of whom (patients 4, 13, and 14) the fusion gene was duplicated (Fig. 1). Patient 4 had one metaphase with an apparent trisomy 21 and two fusion genes; after the FISH analysis the extra chromosome 21 was interpreted as a derivative of the t(12;21)(p13;q22), an observation encountered by others [12,16,17]. All patients with the cryptic t(12;21)(p13;q22) had pre-B ALL. The age at diagnosis of the patients with the fusion gene TEL/AML1 was between 2.5 and 4 years, and all of them attained remission. In two patients, there was an extra signal of the AML1 cosmid probe with no fusion gene. In these patients, the extra signals represented a state of hyperdiploidy that was not detected in the metaphases analyzed by conventional cytogenetics. In patient 1 with the t(12;17)(p13;q12), the TEL signal remained on chromosome 12 indicating that in this
translocation it is not the rearrangement of the TEL gene that plays a role in the malignant transformation. In some of the patients, the proportion of cells with the TEL/ AML1 fusion gene in metaphases was markedly lower than in interphase nuclei. For example, in patient 4, 2 of 25 metaphases had double signals of the fusion gene; in interphase nuclei it was 30 of 50, and with conventional cytogenetic analysis 1 of 25 cells had the der(21)t(12;21)(p13;q22). In patient 6 the TEL/AML1 fusion gene was noted in 4 of 11 metaphases and in 35 of 50 interphase nuclei, and in patient 13, 2 of 22 cells had the TEL/AML1 fusion gene and in the interphase nuclei it was on 91 of 100. This discrepancy is most likely because of the low mitotic index of the leukemic cells under the culture condition, used as part of the cytogenetic analysis. The relatively high incidence (40%) of patients with the t(12;21)(p13;q22) in our study may be the result of the selection method used for the FISH analysis, namely, patients with pre-B ALL that had normal karyotypes at diagnosis. Our study indicates that in childhood B-lineage ALL,
O. Yehuda-Gafni et al. / Cancer Genetics and Cytogenetics 132 (2002) 61–64
63
Table 2 Cytogenetic and fluorescence in situ hybridization results in bone marrow aspirates of the patients at diagnosis and remission Patient no.
Karyotype [no. of cells]
FISH (metaphase) [no. of cells]
FISH (interphase) [no. of cells]
46,XY,t(12;17)(p13;q12) (46%)/46,XY(54%) [24] 46,XY [15] 46,XX [20]
Normal [8]
Normal [50]
Normal [4] 2XTEL, 3XAML1 (22%)/normal (78%) [9]
4
47,XX,21(4%)/46,XX (96%) [25]
5
46,XY [16]
(TEL/AML1)2 (8%)/normal(92%) [25] 2XTEL, 3XAML2 (40%)/normal (60%) [10]
6
46,XX [18]
7
No metaphases
8 9 10 11 12
46,XX [21] 46,XY [10] 46,XY [20] 46,XY [20] No metaphases
Normal [15] Normal [6] Normal [6] Normal [8] TEL/AML1 [12] Remission: normal [25]
13
46,XY [24]
14
No metaphases
(TEL/AML1)2 (9%)/normal (91%) [22] (TEL/AML1)2 [1] Remission: normal [20]
15
46,XX [16]
Normal [50] 2XTEL, 3XAML1 (20%)/normal (80%) [50] Hyperploidy (TEL/AML1)2 60%)/normal (40%) [50] 2XTEL, 3XAML1 (62%)/normal (38%) [50] Hyperploidy TEL/AML1 (70%)/ normal (30%) [50] TEL/AML1 (98%)/ normal (2%) [50] Remission: normal (98%)/ Tel/AML1 (2%) [100] Normal [50] Normal [50] Normal [50] Normal [50] TEL/AML1 (98%)/ normal (2%) [50] Remission: normal [100] (TEL/AML1) 2 (91%)/normal (9%) [100] (TEL/AML1) 2 (94%)/normal (6%) [50] Remission: normal (99%)/TEL/AML1 (1%) [100] Normal [50]
1 2 3
mainly in pre-B ALL with normal karyotypes the cryptic t(12;21)(p13;q22) may account for a high proportion of patients, and therefore FISH analysis is warranted with special attention to interphase nuclei.
References [1] Romana SP, Poirel H, Leconiat M, Flexor MA, Mauchauffe M, Jonveaux P, Macintyre EA, Berger R, Bernard OA. High frequency of t(12;21) in childhood B-lineage acute lymphoblastic leukemia. Blood 1995;86:4263–9. [2] Golub TR, Barker GF, Bohlander SK, Hiebert SW, Ward DC, BrayWard P, Morgan E, Raimondi SC, Rowley JD, Gilliland GD. Fusion of the TEL gene on 12p13 to the AML1 gene on 21q22 in acute lymphoblastic leukemia. Proc Natl Acad Sci USA 1995;92:4917–21. [3] Romana SP, Le Coniat M, Berger R. t(12;21): a new recurrent translocation in acute lymphoblastic leukemia. Genes Chromosomes Cancer 1994;9:186–91.
TEL/AML1 (36%)/ normal (64%) [11] TEL/AML1 [2] Remission: normal [15]
Normal [16]
[4] Amor DJ, Algar EM, Slater HR, Smith PJ. High frequency of t(12;21) in childhood acute lymphoblastic leukemia detected by RT-PCR. Pathology 1998;30:381–5. [5] Rubnitz JE, Downing JR, Pui CH, Shurtleff SA, Raimondi SC, Evans WE, Head DR, Crist WM, Rivera GK, Hancock ML, Boyett JM, Buijs A, Grosveld G, Behm FG. TEL gene rearrangement in acute lymphoblastic leukemia: a new genetic marker with prognostic significance. J Clin Oncol 1997;15:1150–7. [6] Shurtleff SA, Buijs A, Behm FG, Rubnitz JE, Raimondi SC, Hancock ML, Chan GC, Pui CH, Grosveld G, Downing JR. TEL/AML1 fusion resulting from a cryptic t(12;21) is the most common genetic lesion in pediatric ALL and defines a subgroup of patients with an excellent prognosis. Leukemia 1995;9:1985–9. [7] Liang DC, Chou TB, Chen JS, Shurtleff SA, Rubnitz JE, Downing JR, Pui CH, Shih LY. High incidence of TEL/AML1 fusion resulting from a cryptic t(12;21) in childhood B-lineage acute lymphoblastic leukemia in Taiwan. Leukemia 1996;10:991–3. [8] McLean TW, Ringold S, Neuberg D, Stegmaier K, Tantravahi R, Ritz J, Koeffler HP, Takeuchi S, Janssen JW, Seriu T, Bartram CR, Sallan SE, Gilliland DG, Golub TR. TEL/AML-1 dimerizes and is associ-
64
O. Yehuda-Gafni et al. / Cancer Genetics and Cytogenetics 132 (2002) 61–64
[9]
[10]
[11]
[12]
[13]
[14]
[15]
[16] [17]
Fig. 1. Fluorescence in situ hybridization analysis of patient 13 using TEL (green) and AML1 (red) cosmid probes. (a) Cells with no t(12;21)(p13; q22). (b) Cells with t(12;21)(p13;q22); two der(21)t(12;21)(p13;q22) chromosomes (arrowheads). The arrows points to the residual signal of the AML1 cosmid probe on the derivative chromosome 12.
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