Premium
Application of laser capture microdissection to cytologic specimens for the detection of immunoglobulin heavy chain gene rearrangement in patients with malignant lymphoma
Author(s) -
Orba Yasuko,
Tanaka Shinya,
Nishihara Hiroshi,
Kawamura Naoki,
Itoh Tomoo,
Shimizu Michio,
Sawa Hirofumi,
Nagashima Kazuo
Publication year - 2003
Publication title -
cancer cytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.052
H-Index - 304
eISSN - 1097-0142
pISSN - 0008-543X
DOI - 10.1002/cncr.11331
Subject(s) - gene rearrangement , laser capture microdissection , lymphoma , biology , polymerase chain reaction , pathology , immunoglobulin heavy chain , microdissection , population , microbiology and biotechnology , cytology , immunoglobulin gene , genomic dna , single strand conformation polymorphism , antibody , gene , medicine , genetics , gene expression , environmental health
BACKGROUND The demonstration of the monoclonality of immunoglobulin heavy chain (IgH) gene rearrangement is an indispensable method for the diagnosis of B‐cell lymphoma as well as histocytochemical analysis. For the detection of IgH gene rearrangement, the extraction of DNA from a homogenous cell population is necessary. Recently, the laser capture microdissection (LCM) technique was shown to isolate specific cells from histopathologic specimens for molecular analysis. However, to the authors' knowledge the applicability of LCM to cytologic specimens has not yet been well established. METHODS Using LCM, a homogenous population of B‐cell lymphoma cells as both histologic sections and cytologic specimens was captured, and genomic DNA was extracted from the captured cells. IgH gene rearrangement was analyzed by the polymerase chain reaction (PCR)‐based single‐strand conformational polymorphism (SSCP) method. RESULTS Genomic DNAs were extracted successfully from ethanol‐fixed cytologic specimens, but cells were not captured from air‐dried specimens. Using PCR‐SSCP analysis, the monoclonality of the IgH gene rearrangement was detected in five cases of tissue sections among nine analyzed cases of malignant lymphoma diagnosed immunohistochemically. However, analysis of the cytologic specimens with LCM demonstrated the monoclonality of the IgH gene rearrangement in seven cases of lymphoma. CONCLUSIONS The results of the current study suggest that the novel application of LCM to cytologic specimens occasionally exhibits high sensitivity for the detection of IgH gene rearrangement monoclonality compared with the use of histologic sections. Cancer (Cancer Cytopathol) 2003;99:198–204. © 2003 American Cancer Society.