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Detection of Trisomy 8 in Philadelphia Chromosome–Positive CML Patients Using Conventional Cytogenetic and Interphase Fluorescence in situ Hybridization Techniques and its Relation to c-myc Involvement

Address correspondence to Armand B. Glassman, M.D., The University of Texas, M. D. Anderson Cancer Center, 1515 Holcombe Blvd, Box 350, Houston, Texas 77030-4095, USA; tel 713 792 6330; fax 713 745 3215; email acglassman{at}juno.com.

Trisomy 8 (+8) is a common clonal evolution marker for progression in chronic myelogenous leukemia. The relationship of +8 to various stages of t(9;22) leukemias is not firmly established. To explore this association we examined bone marrow (BM) cells from 10 Philadelphia chromosome positive (Ph+) chronic myeloid leukemia (CML) patients in different stages of the disease, using conventional cytogenetic technique(CCT) and interphase fluorescence in situ hybridization (FISH). FISH detection of chromosome 8 was accomplished using the D8Z2 (Oncor) probe specific for the centrometric region of chromosome 8. Five hundred interphase nuclei were counted for each patient. Three of the 10 patients were selected for detection of c-myc gene locus located in the 8q24.2–24.3 region using the LSI c-myc probe (Vysis). One hundred interphase nuclei were counted for the presence of the c-myc gene for each patient. We found that the percentage of interphase nuclei showing 3 hybridization spots indicative of trisomy 8 was significantly lower by FISH than by CCT (metaphase analysis) in patients with chronic phase (CP) CML (11% versus 24%), nearly similar in patients with accelerated phase (AP) CML (13% versus 10%), and significantly higher in patients with myeloid blast crisis (mBC) (81% versus 33%). Hybridization spots for the c-myc locus were consistent with the chromosome 8 interphase FISH results in each of the patients tested. It is hypothesized that cells with supernumerary chromosome 8 may have a cell cycle time that differs from that of the disomic cells according to the stage of the disease. The c-myc locus expression in Ph+ CML patients with trisomy 8 is related to an increased copy number of the gene.

Keywords: chronic myeloid leukemia, CML, c-myc. trisomy 8, Philadelphia chromosome, FISH