Genomic analysis of CD8+ NK/T cell line, 'SRIK-NKL', with array-based CGH (aCGH), SKY/FISH and molecular mapping.
We performed aCGH, SKY/FISH, molecular mapping and expression analyses on a permanent CD8+ NK/T cell line, 'SRIK-NKL' established from a lymphoma (ALL) patient, in attempt to define the fundamental genetic profile of its unique NK phenotypes. aCGH revealed hemizygous deletion of 6p containing genes responsible for hematopoietic functions. The SKY demonstrated that a constitutive reciprocal translocation, rcpt(5;14)(p13.2;q11) is a stable marker. Using somatic hybrids containing der(5) derived from SRIK-NKL, we found that the breakpoint in one homologue of no. 5 is located upstream of IL7R and also that the breakpoint in no. 14 is located within TRA@. The FISH analysis using a BAC which contains TRA@ and its flanking region further revealed a approximately 231kb deletion within 14q11 in the der(5) but not in the normal homologue of no. 14. The RT-PCR analysis detected mRNA for TRA@ transcripts which were extending across, but not including, the deleted region. IL7R was detected at least at mRNA levels. These findings were consistent with the immunological findings that TRA@ and IL7R are both expressed at mRNA levels and TRA@ at cytoplasmic protein levels in SRIK-NKL cells. In addition to rcpt(5;14), aCGH identified novel copy number abnormalities suggesting that the unique phenotype of the SRIK-NKL cell line is not solely due to the TRA@ rearrangement. These findings provide supportive evidence for the notion that SRIK-NKL cells may be useful for studying not only the function of NK cells but also genetic deregulations associated with leukemiogenesis.
Rossi
Michael R
MR
SKY Core Facility, Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, NY 14263, United States.
Laduca
Jeff
J
Cowell
John K
JK
Srivastava
Bejai I S
BI
Matsui
Seiichi
S
D016827:CD8 Antigens / Q000378:metabolism*; D002460:Cell Line; D019457:Chromosome Breakage; D002874:Chromosome Mapping; D002883:Chromosomes, Human, Pair 14; D002895:Chromosomes, Human, Pair 5; D020732:Cytogenetic Analysis; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007694:Killer Cells, Natural / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D032681:Spectral Karyotyping; D013601:T-Lymphocytes / Q000378:metabolism*; D014178:Translocation, Genetic
D016827:CD8 Antigens