PMC7441545_01
Cytogenetic analysis of bone marrow cells at diagnosis of T-NHL in 6-year-old male. (a) The karyogram (GTG-banding) showing complex karyotype of the patient: 45,XY,-1, dup(1)(p32p34),der(3)t(1;3)(q12;q22),der(5)t(5;10)(q21;p11),der(9)t(4;9)(q32;p21),der(11)t(1;11)(p32;p13),del(12)(p13),der(16)t(6;16)(q22;p13)[10]/46,XY[15] (b,c,d) Results of FISH tests with probes: ETV6/RUNX1, BCR/ABL1 and KMT2A. FISH was performed on metaphases and interphase nuclei using probes (Cytocell Ltd., Oxford Gene Technology, Cambridge, United Kingdom) according to the manufacturer's recommendations. Images were captured by an Olympus BX41TF microscope equipped with a Jenoptik camera and analysed with Isis Software (MetaSystems)
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Fig1
GTG band staining (Fig. 1a) and fluorescence in situ hybridization (FISH) test were performed with the use of probes: BCR/ABL1, KMT2A, ETV6/RUNX1 (Vysis, Abbot Molecular, Illinois, USA). The arrangement from ETV6/RUNX1 probe suggested ETV6 deletion (Fig. 1b). The arrangement of signals from other probes used was correct (Fig. 1c and d). FISH tests (Fig. 3). Finally cytogenetic result was the following: 45,XY,-1,dup(1)(p32p34),der(3)t(1;3)(q12;q22),der(5)t(5;10)(q21;p11),der(9)t(4;9)(q32;p21),der(11)t(1;11)(p32;p13),del(12)(p13),der(16)t(6;16)(q22;p13)[10]/46,XY[15] (Fig. 4 and Table 1).
Karyoview from microarray test and a scheme presenting chromosomal aberrations in patient. Microarray results revealing partial gains of overlapping regions on chromosomes 1p 8,931,529-67,365,806 bp (1p36.23-p31.3), 4q 155,500,158-190,957,473 bp (4q31.3-q35.2), 6q 115,144,178-170,919,482 bp (6q22.1-q27) and 10p 100,026-38,258,848 bp (10p15.3-p11.1). Moreover, regions of overlap of deletions were also found on chromosomes 1p 849,466-8,096,240 bp (1p36.33-p36.23) and 1 70,493,564-145,289,186 bp (1p31.12-q21.1), 5q 100,821,228-180,719,789 bp (5q21.1-q35.3), 9p 203,861-28,849,504 bp (9p24.3-p21.1), 11p 230,615-35,363,338 bp (11p15.5-p13), 12p 173,786-22,885,159 bp (12p13.33-p12.1) and 16p 85,880-10,023,421 bp (16p13.3-p13.2). Asterisks correspond to deletion (red colour), duplication (blue colour) and loss of heterozygosity (purple colour)
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Fig2
Thermo Fischer Scientific, Waltham, MA, USA). Tests revealed additional alterations in the form of gained copies (4q32-q35, 6q22-q27, 10p11-p15) and loss regions (9p21-p24, 5q21-q35) (Fig. 2).
Images of the FISH results revealing chromosomal aberrations. (a) Image of the FISH results with the whole chromosome painting (WCP) 1 and 3 probes (Cytocell Ltd., Oxford Gene Technology, Cambridge, United Kingdom) revealing t(1;3). (b) Chromosome analysis demonstrating derivative chromosomes 3 der(3)t(1;3)(q12;q22) and chromosome 1. (c) Image of the FISH results with the LSI CSF1R/D5S23, D5S721 Dual Color probe (Vysis, Abbot Molecular, Illinois, USA) revealing del(5q33-q34). (d) Chromosome analysis demonstrating abnormal chromosome 5 with deletion of 5q33-q34. (e) Image of the FISH results with the STIL Break Apart Probe (Empire Genomics, New York, USA) revealing STIL duplication. (f) Image of the FISH results with WCP4 probe (Cytocell Ltd., Oxford Gene Technology, Cambridge, United Kingdom) revealing t(4;9). (g) Image of the FISH results with WCP5 and WCP10 probes (Cytocell Ltd., Oxford Gene Technology, Cambridge, United Kingdom) revealing t(5;10). (h) Image of the FISH results with WCP6 and WCP16 probes (Cytocell Ltd., Oxford Gene Technology, Cambridge, United Kingdom) revealing t(6;16). Images were captured by an Olympus BX41TF microscope equipped with a Jenoptik camera and analysed with Isis Software (MetaSystems)
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Fig3
FISH tests (Fig. 3). Finally cytogenetic result was the following: 45,XY,-1,dup(1)(p32p34),der(3)t(1;3)(q12;q22),der(5)t(5;10)(q21;p11),der(9)t(4;9)(q32;p21),der(11)t(1;11)(p32;p13),del(12)(p13),der(16)t(6;16)(q22;p13)[10]/46,XY[15] (Fig. 4 and Table 1).
The scheme of chromosomal aberrations in patient based on cytogenetic analysis, microarray tests and FISH results prepared using CyDAS software (http://www.cydas.org/OnlineAnalysis/, Duesseldorff, Germany). Hash represents derivative chromosomes
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Fig4
FISH tests (Fig. 3). Finally cytogenetic result was the following: 45,XY,-1,dup(1)(p32p34),der(3)t(1;3)(q12;q22),der(5)t(5;10)(q21;p11),der(9)t(4;9)(q32;p21),der(11)t(1;11)(p32;p13),del(12)(p13),der(16)t(6;16)(q22;p13)[10]/46,XY[15] (Fig. 4 and Table 1).