Genetic analysis of sorted Hodgkin and Reed-Sternberg cells using comparative genomic hybridization.
Hodgkin and Reed-Sternberg (H and RS) cells are generally considered to be the neoplastic cells of Hodgkin's disease (HD); however, such cells are found only in a minority of HD lesions. Very few data have so far been published on the cytogenetic abnormalities in HD. An analysis of unknown genetic aberrations has only recently become possible through the use of comparative genomic hybridization (CGH), which is based on the competitive binding of tumor and control DNA to metaphase chromosomes. In order to exclude the reaction of non-tumor cells, we used 100 sorted H-RS cells as the tumor DNA, then 100 sorted reactive T cells or B cells as the control DNA. We obtained the amplified DNA, using degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). In addition, to confirm whether or not the lymphocytes in the background were reactive, we performed CGH with 100 sorted B cells and 100 sorted T cells. CGH was thus performed on 9 HDs, including 6 cases of mixed-cellularity (MC) sub-type and 3 cases of nodular-sclerosis (NS) sub-type. CGH of the B and T cells showed no genetic changes in any cases. In contrast, CGH of H-RS cells revealed both gains and losses of DNA in all 9 cases, and multiple changes were also observed. In situ hybridization showed an Epstein-Barr-virus infection in 5 cases of MC; however, no definite relationship was observed between the EBV infection and genetic changes. The most commonly observed genetic aberrations were a loss on 16q11/21 in 6 cases, a gain on 1p13 in 5 cases, and a gain on 7q35/36 in 5 cases. The large number of chromosomal alterations in HD suggests, therefore, that an increased degree of genetic instability play a role in the formation of H-RS cells.
Ohshima
K
K
Department of Pathology, School of Medicine, Fukuoka University, Japan.
Ishiguro
M
M
Ohgami
A
A
Sugihara
M
M
Haraoka
S
S
Suzumiya
J
J
Kikuchi
M
M
D000328:Adult; D000368:Aged; D001402:B-Lymphocytes / Q000737:chemistry; D002469:Cell Separation; D002648:Child; D002869:Chromosome Aberrations; D002885:Chromosomes, Human, Pair 16 / Q000235:genetics; D002999:Clone Cells / Q000737:chemistry / Q000473:pathology; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D020031:Epstein-Barr Virus Infections / Q000235:genetics / Q000473:pathology*; D005260:Female; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006689:Hodgkin Disease / Q000235:genetics / Q000473:pathology* / Q000821:virology; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D008875:Middle Aged; D009154:Mutation; D014411:Neoplastic Stem Cells / Q000737:chemistry* / Q000821:virology; D016133:Polymerase Chain Reaction; D016539:Reed-Sternberg Cells / Q000737:chemistry* / Q000821:virology; D017384:Sequence Deletion; D013601:T-Lymphocytes / Q000737:chemistry
D017931:DNA Primers; D004273:DNA, Neoplasm; D004279:DNA, Viral