Detection of t(4;14)(p16.3;q32) Chromosomal Translocation in Multiple Myeloma by Double-Color Fluorescent In Situ Hybridization

Abstract

Chromosomal translocations involving the immunoglobulin heavy chain (IGH) locus at chromosome 14q32 represent a common mechanism of oncogene activation in lymphoid malignancies. In multiple myeloma (MM), variable chromosome partners have been identified by conventional cytogenetics, including the 11q13, 8q24, 18q21, and 6p21 loci. We and others have recently reported a novel, karyotypically undetectable chromosomal translocation t(4;14)(p16.3;q32) in MM-derived cell lines, as well as in primary tumors. The 4p16.3 breakpoints are relatively scattered and located less than 100 kb centromeric of the fibroblast growth factor receptor 3 (FGFR3) gene or within the recently identified WHSC1 gene, both of which are apparently deregulated by the translocation. To assess the frequency of the t(4;14)(p16.3;q32) translocation in MM, we performed a double-color fluorescent in situ hybridization (FISH) analysis of interphase nuclei with differently labeled probes specific for the IGH locus (a pool of plasmid clones specific for the IGH constant regions) or 4p16.3 (yeast artificial chromosome (YAC) 764-H1 spanning the region involved in breakpoints). Thirty MM patients, the MM-derived cell lines KMS-11 and OPM2, and six normal controls were examined. The identification of a t(4;14) translocation, evaluated as the presence of a der(14) chromosome, was based on the colocalization of signals specific for the two probes; a cutoff value of 15% (mean + 3 standard deviation [SD]) derived from the interphase FISH of the normal controls (range, 5% to 11%; mean ± SD, 8.16 ± 2.2) was used for the quantification analysis. In interphase FISH, five patients (one in clinical stage I, two in stage II, one in stage III, and a plasma cell leukemia) were found to be positive (≈15%). FISH metaphases with split or colocalized signals were detected in only two of the translocated cases and confirmed the pattern found in the interphase nuclei. Furthermore, in three of the five cases with the translocation, FISH analysis with the IGH joining probe (JH) showed the presence of the reciprocal product of the translocation [der(4) chromosome]. Overall, our study indicates that the t(4;14)(p16.3;q32) chromosomal translocation is a recurrent event in MM tumors and may contribute towards the detection of this lesion and our understanding of its pathogenetic and clinical implications in MM.

Functional regulation of D‐type cyclins by insulin‐like growth factor‐I and serum in multiple myeloma cells

“…KMS12‐BM, KMS12‐PE, U266 and XG1 all carried the 11q13 translocation, t(11;14), which leads to the expression of CCND1 , (Zhang et al , 1994; Vaandrager et al , 1997; Gabrea et al , 1999) and all were positive for cyclin D1 protein. Of the cyclin D2 positive HMCLs, MM‐1S, JJN3 and RPMI‐8226 carried the t(14;16) translocation that leads to overexpression of CCND2 , via c‐maf (Chesi et al , 1998), while NCI‐H929 and OPM2 were positive for MMSET via a t(4;14) which also leads to CCND2 overexpression (Finelli et al , 1999; Grand et al , 2004). Interestingly, KMS28‐BM/PE are positive for t(4;14), but express cyclin D3 instead of cyclin D2, consistent with previous mRNA expression profiling (Lombardi et al , 2007).…”

Malignant hematological disorders in children with Wolf–Hirschhorn syndrome

“…Hence, it is possible that either WHSC1 and/or FGFR3 gene act as tumor suppressor gene and when completely disrupted or deleted as in our cases, disturb the normal hematopoiesis and lead to malignant transformation. It is noteworthy that in another tumor model, multiple myeloma carrying the translocation t(4;14)(p16.3;q32), both these genes are fused with IgH gene [Finelli et al, 1999; Nakazawa et al, 2000]. In contrast to the carcinoma model mentioned above, in the multiple myeloma model the resultant fusion transcript produces an abnormal chimeric protein, which is over‐expressed and is responsible for malignant hematopoietic progression.…”

Immunohistochemical Analysis of Cyclin D1 Shows Deregulated Expression in Multiple Myeloma with the t(11;14)

“…FISH analyses were possible on 39 of the 48 patient samples and were performed as previously described. 11 The percentage of malignant plasma cells, assessed by morphologically analyzing cytospins of cell suspensions from bone marrows, ranged from 13 to 85% (median, 31%). The bone marrow cells were cultured in RPMI 1640 medium supplemented with 20% fetal calf serum without any mitogen for 24 hours at 37°C in 5% CO 2 .…”