scholarly journals Detection of the Philadelphia chromosome in acute lymphoblastic leukemia by pulsed-field gel electrophoresis

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 1101-1107 ◽  
Author(s):  
AL Hooberman ◽  
CM Rubin ◽  
KP Barton ◽  
CA Westbrook
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Gel Electrophoresis ◽  
Chromosomal Abnormalities ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
Pulsed Field Gel Electrophoresis ◽  
Myelogenous Leukemia ◽  
Clinical Samples ◽  
Pulsed Field ◽  
Translocation Breakpoints

Abstract The Philadelphia (Ph1) chromosome is an acquired abnormality in the malignant cells of 10% to 25% of patients with acute lymphoblastic leukemia (ALL). Unlike chronic myelogenous leukemia (CML), where the molecular detection of the Ph1 chromosome is relatively straightforward using conventional Southern hybridization analysis, the detection of the Ph1 chromosome in ALL is complicated by the existence of several molecular subtypes, and the fact that translocation breakpoints are dispersed over a large genomic area. To circumvent these difficulties, we investigated pulsed-field gel electrophoresis (PFGE) to determine if this method could be used directly on clinical samples to detect the Ph1 chromosome in ALL. We report that, in a study of seven patients with Ph1-positive ALL, we could easily detect the Ph1 using only a single PFGE analysis, regardless of the Ph1 subtype, and we could confirm that the translocations occur either within or very near the BCR gene in all seven. We conclude that PFGE is a useful technique for the detection of the Ph1 in ALL, which ultimately may find wide applicability in the detection of other chromosomal abnormalities in other malignancies.

scholarly journals Detection of the Philadelphia chromosome in acute lymphoblastic leukemia by pulsed-field gel electrophoresis

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 1101-1107
Author(s):  
AL Hooberman ◽  
CM Rubin ◽  
KP Barton ◽  
CA Westbrook
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Gel Electrophoresis ◽  
Chromosomal Abnormalities ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
Pulsed Field Gel Electrophoresis ◽  
Myelogenous Leukemia ◽  
Clinical Samples ◽  
Pulsed Field ◽  
Translocation Breakpoints

The Philadelphia (Ph1) chromosome is an acquired abnormality in the malignant cells of 10% to 25% of patients with acute lymphoblastic leukemia (ALL). Unlike chronic myelogenous leukemia (CML), where the molecular detection of the Ph1 chromosome is relatively straightforward using conventional Southern hybridization analysis, the detection of the Ph1 chromosome in ALL is complicated by the existence of several molecular subtypes, and the fact that translocation breakpoints are dispersed over a large genomic area. To circumvent these difficulties, we investigated pulsed-field gel electrophoresis (PFGE) to determine if this method could be used directly on clinical samples to detect the Ph1 chromosome in ALL. We report that, in a study of seven patients with Ph1-positive ALL, we could easily detect the Ph1 using only a single PFGE analysis, regardless of the Ph1 subtype, and we could confirm that the translocations occur either within or very near the BCR gene in all seven. We conclude that PFGE is a useful technique for the detection of the Ph1 in ALL, which ultimately may find wide applicability in the detection of other chromosomal abnormalities in other malignancies.

scholarly journals Long-range mapping of the Philadelphia chromosome by pulsed-field gel electrophoresis

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 697-702
Author(s):  
CA Westbrook ◽  
CM Rubin ◽  
JJ Carrino ◽  
MM Le Beau ◽  
A Bernards ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Large Scale ◽  
Fusion Gene ◽  
Philadelphia Chromosome ◽  
Pulsed Field Gel Electrophoresis ◽  
Chromosome 9 ◽  
Myelogenous Leukemia ◽  
Chromosome 22 ◽  
Pulsed Field ◽  
Translocation Breakpoints

The Philadelphia chromosome (Ph1) of chronic myelogenous leukemia (CML) contains sequences from chromosome 9, including the ABL protooncogene, that have been translocated to the breakpoint cluster region (bcr) of chromosome 22, giving rise to a bcr-ABL fusion gene, whose product has been implicated in the genesis of CML. Although chromosome 22 translocation breakpoints in CML virtually always occur within the 5.8- kilobase (kb) bcr, chromosome 9 breakpoints have been identified within the known limits of ABL in only a few instances. For a better understanding of the variability of the breakpoints on chromosome 9, we studied the CML cell line BV173. Using pulsed-field gel electrophoresis (PFGE), large-scale maps of the t(9;22) junctions were constructed. The chromosome 9 breakpoint was shown to have occurred within an ABL intron, 160 kb upstream of the v-abl homologous sequences, but still 35 kb downstream of the 5′-most ABL exon. bcr-ABL and ABL-bcr fusion genes were demonstrated on the Ph1 and the 9q+ chromosomes, respectively; both of these genes are expressed. These results suggest that the 9;22 translocation breakpoints in CML consistently occur within the limits of the large ABL gene. RNA splicing, sometimes of very large regions, appears to compensate for the variability in breakpoint location. These studies show that PFGE is a powerful new tool for the analysis of chromosomal translocations in human malignancies.

scholarly journals Long-range mapping of the Philadelphia chromosome by pulsed-field gel electrophoresis

Blood ◽  
1988 ◽  
Vol 71 (3) ◽  
pp. 697-702 ◽  
Author(s):  
CA Westbrook ◽  
CM Rubin ◽  
JJ Carrino ◽  
MM Le Beau ◽  
A Bernards ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Large Scale ◽  
Fusion Gene ◽  
Philadelphia Chromosome ◽  
Pulsed Field Gel Electrophoresis ◽  
Chromosome 9 ◽  
Myelogenous Leukemia ◽  
Chromosome 22 ◽  
Pulsed Field ◽  
Translocation Breakpoints

Abstract The Philadelphia chromosome (Ph1) of chronic myelogenous leukemia (CML) contains sequences from chromosome 9, including the ABL protooncogene, that have been translocated to the breakpoint cluster region (bcr) of chromosome 22, giving rise to a bcr-ABL fusion gene, whose product has been implicated in the genesis of CML. Although chromosome 22 translocation breakpoints in CML virtually always occur within the 5.8- kilobase (kb) bcr, chromosome 9 breakpoints have been identified within the known limits of ABL in only a few instances. For a better understanding of the variability of the breakpoints on chromosome 9, we studied the CML cell line BV173. Using pulsed-field gel electrophoresis (PFGE), large-scale maps of the t(9;22) junctions were constructed. The chromosome 9 breakpoint was shown to have occurred within an ABL intron, 160 kb upstream of the v-abl homologous sequences, but still 35 kb downstream of the 5′-most ABL exon. bcr-ABL and ABL-bcr fusion genes were demonstrated on the Ph1 and the 9q+ chromosomes, respectively; both of these genes are expressed. These results suggest that the 9;22 translocation breakpoints in CML consistently occur within the limits of the large ABL gene. RNA splicing, sometimes of very large regions, appears to compensate for the variability in breakpoint location. These studies show that PFGE is a powerful new tool for the analysis of chromosomal translocations in human malignancies.

Philadelphia chromosome and monosomy 7 in childhood acute lymphoblastic leukemia: a Pediatric Oncology Group study

Blood ◽  
1991 ◽  
Vol 77 (5) ◽  
pp. 1050-1056 ◽  
Author(s):  
C Russo ◽  
A Carroll ◽  
S Kohler ◽  
M Borowitz ◽  
M Amylon ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukemia ◽  
Chronic Myelogenous Leukemia ◽  
Pediatric Oncology ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
Myelogenous Leukemia ◽  
Oncology Group ◽  
Monosomy 7 ◽  
Pediatric Oncology Group

Abstract During an 8-year period, 3,638 children from institutions of the Pediatric Oncology Group (POG) were diagnosed with acute lymphoblastic leukemia (ALL). Fifty-seven patients had Philadelphia chromosome- positive (Ph1) ALL. Blast cells obtained at diagnosis from 13 of these 57 cases (23%) were also found to have partial or complete monosomy 7 (- 7). This subgroup of children with Ph1/-7 ALL was comprised primarily of older males with early B-lineage ALL. Bone marrow specimens from six Ph1/-7 patients were studied further using the polymerase chain reaction and primers that flank the ALL, and chronic myelogenous leukemia breakpoints to determine the molecular characteristic of the 9;22 translocation. Rearrangements were detected in RNA from bone marrow and/or peripheral blood cells of six patients, although four were in hematologic remission at the time of the analysis. Five cases showed the ALL breakpoint, while one child with Ph1/-7 showed the chronic myelogenous leukemia breakpoint. The induction failure rate was much higher in this subgroup (31%) as compared with Ph1-negative cases, and the projected duration of event-free survival reflected the aggressive nature of this subgroup because no children are projected to remain in remission at 2 years. ALL with both the 9;22 translocation and -7 appears to represent a unique and previously undescribed subgroup of childhood ALL associated with a particularly adverse outcome. Leukemic transformation in such patients may involve the interaction of a dominant oncogene (Ph1) and a tumor suppressor gene (- 7).

Sign in / Sign up

Export Citation Format

Share Document