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Published on: February 3, 2013
Interphase cytogenetics of non-Hodgkin's lymphoma using non-fluorescent in situ hybridization in paraffin embedded
1Department of Clinical Pathology and Pediatrics, Catholic University Medical College, Seoul, Korea.
Insights
Non-Hodgkin's lymphoma (NHL) shows frequent chromosomal aberrations, particularly extracopy of the X chromosome. Specific aberrations correlate with NHL grades, aiding diagnosis and outcome assessment.
Area of Science:
- Cytogenetics
- Oncology
- Molecular Pathology
Background:
- Non-Hodgkin's lymphoma (NHL) is a heterogeneous group of lymphoid malignancies.
- Accurate diagnosis and prognostication are crucial for effective NHL management.
- Interphase cytogenetic studies can reveal chromosomal abnormalities in archival tissues.
Purpose of the Study:
- To investigate chromosomal aberrations in paraffin-embedded NHL tissues using non-fluorescent in situ hybridization (NFISH).
- To correlate specific chromosomal aberrations with histopathologic grades and subtypes of NHL.
- To evaluate the utility of interphase NFISH for retrospective cytogenetic analysis.
Main Methods:
- Paraffin-embedded tissue samples from 30 NHL cases and 10 reactive hyperplasia controls were analyzed.
- Interphase non-fluorescent in situ hybridization (NFISH) was performed using enzymatic methods.
- Digoxigenin-labeled DNA probes for chromosomes 7, 12, 18, X, and a chromosome 18 painting probe were utilized.
Main Results:
- Chromosomal aberrations were detected in 90% (27/30) of NHL cases.
- Extracopy of the X chromosome was the most common numerical aberration.
- Specific aberrations correlated with NHL grades: extracopy of chromosome 12 in low-grade NHL, and tetraploidy and chromosome 18 aberrations in intermediate-grade NHL.
Conclusions:
- Interphase NFISH is a feasible method for cytogenetic studies on archival tissue samples.
- Specific chromosomal aberrations are associated with NHL histopathologic grades and subtypes.
- These findings can enhance pathologic diagnosis and clinical outcome assessment for NHL patients.
Abstract:
Paraffin-embedded tissue samples from 30 cases of non-Hodgkin's lymphoma(NHL) and 10 of reactive hyperplasia, were processed for interphase cytogenetic chromosomal study. We performed non-fluorescent in situ hybridization(NFISH) using the enzymatic method with digoxigenin-labeled DNA centromeric probes for chromosome 7,12,18 and X, and a painting probe for chromosome 18. Chromosomal aberrations were observed in 27(90%) out of 30 cases of NHL. The most commonly observed numerical aberration was extracopy of X chromosome. There were some characteristic aberrations corresponding to each grade and group of NHL by International Working Formulation: In low grade NHL(9 cases), a third were associated with extracopy of chromosome 12, and disomy X was frequently found in small lymphocytic lymphoma(75%). With intermediate grade(16 cases), tetraploidy(25%), translocation of chromosome 18(25%), and extracopy of chromosome 18(19%) were characteristically associated. These results suggest that interphase NFISH is an easily performable method in retrograde cytogenetic study of archival materials. Some specifically correlated chromosomal aberrations corresponding to the histopathologic grades and groups could provide us more valuable information for determining pathologic diagnosis and assessing the clinical outcome of NHL.
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