Multicolour-banding fluorescence in situ hybridisation (mbanding-FISH) to identify recurrent chromosomal alterations in breast tumour cell lines

Abstract

Recurrent chromosome breakpoints in tumour cells may point to cancer genes, but not many have been molecularly characterised. We have used multicolour-banding fluorescence in situ hybridisation (mbanding-FISH) on breast tumour cell lines to identify regions of chromosome break created by inversions, duplications, insertions and translocations on chromosomes 1, 5, 8, 12 and 17. We delineate a total of 136 regions of break, some of them occurring with high frequency. We further describe two examples of dualcolour FISH characterisation of breakpoints, which target the 1p36 and 5p11 -12 regions. Both breaks involve genes whose function is unknown to date. The mbanding-FISH strategy constitutes an efficient first step in the search for potential cancer genes.

Comprehensive Profiling of 8p11-12 Amplification in Breast Cancer

“…Regions were considered to be gained or lost when at least two consecutive BAC clones presented a log 2 ratio > 0. 25 classified by hierarchical clustering (Fig. 3A).…”

“…We used dual-color FISH analysis with probes covering the region between NRG1 (not included) and FGFR1 ( Fig. 2F; Supplementary Table S4) in a series of 12 cell lines (Set 1, Supplementary Table S1) selected based on either the existence of transition in their aCGH profile or 8p11-21 rearrangement detected previously by mbanding FISH (25).…”

“…BAC clones were checked for chimerism by FISH on normal metaphases chromosomes. After counterstaining with Vectashield containing 4V ,6-diamidino-2-phenylindole (Vector, Burlingame, CA), images were processed as described previously (25).…”

“…Break characterization was based on the split-signal FISH approach (25,57). Tumor sections present on tissue microarray were first hybridized with biotinylated and digoxigenin-labeled BAC combinations P1 + P5 to determine the presence of alterations (breaks and/or amplification) in the [UNC5D-FGFR1] genomic region.…”

Multicolor banding detects a complex three chromosome, seven breakpoint unbalanced rearrangement in an ICSI‐derived fetus with multiple abnormalities

“…Previous publications describing the use of MCB include: diagnosis of a balanced CCR found in a healthy female as part of infertility investigations [Kuechler et al, 2005], prenatal diagnosis of a balanced CCR in an ICSI pregnancy [Trimborn et al, 2005], characterization of a 15‐breakpoint CCR in a mentally retarded child [Houge et al, 2003] and a multiple translocation event in a patient with hexadactyly, facial dysmorphism, mental retardation, and behavior disorder [Seidel et al, 2003], confirmation of a complex karyotype in a child with multiple congenital abnormalities [Kline et al, 2004], and the identification of recurrent chromosomal alterations in breast tumor cell lines [Letessier et al, 2005]. In the case described here, the structure of the derivative chromosome 12, together with the nature and extent of the monosomy, could not have been ascertained without the use of MCB; we are only aware of one other publication [Weise et al, 2002], describing such delineation of chromosomal monosomy in a constitutional karyotype.…”