title: Non-random distribution of instability-associated chromosomal rearrangement breakpoints in human lymphoblastoid cells.
abstract: Genomic instability is observed in tumors and in a large fraction of the progeny surviving irradiation. One of the best-characterized phenotypic manifestations of genomic instability is delayed chromosome aberrations. Our working hypothesis for the current study was that if genomic instability is in part attributable to cis mechanisms, we should observe a non-random distribution of chromosomes or sites involved in instability-associated rearrangements, regardless of radiation quality, dose, or trans factor expression. We report here the karyotypic examination of 296 instability-associated chromosomal rearrangement breaksites (IACRB) from 118 unstable TK6 human B lymphoblast, and isogenic derivative, clones. When we tested whether IACRB were distributed across the chromosomes based on target size, a significant non-random distribution was evident (p<0.00001), and three IACRB hotspots (chromosomes 11, 12, and 22) and one IACRB coldspot (chromosome 2) were identified. Statistical analysis at the chromosomal band-level identified four IACRB hotspots accounting for 20% of all instability-associated breaks, two of which account for over 14% of all IACRB. Further, analysis of independent clones provided evidence within 14 individual clones of IACRB clustering at the chromosomal band level, suggesting a predisposition for further breaks after an initial break at some chromosomal bands. All of these events, independently, or when taken together, were highly unlikely to have occurred by chance (p<0.000001). These IACRB band-level cluster hotspots were observed independent of radiation quality, dose, or cellular p53 status. The non-random distribution of instability-associated chromosomal rearrangements described here significantly differs from the distribution that was observed in a first-division post-irradiation metaphase analysis (p=0.0004). Taken together, these results suggest that genomic instability may be in part driven by chromosomal cis mechanisms.
authors.affiliation: Environmental Toxicology Graduate Program, Department of Cell Biology and Neuroscience, University of California, Riverside, CA, USA.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Moore
authors.forename: David
authors.initials: D
authors.lastname: Papworth
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Grosovsky
mesh_terms: D001402:B-Lymphocytes / Q000648:ultrastructure; D002460:Cell Line; D019457:Chromosome Breakage; D003627:Data Interpretation, Statistical; D042822:Genomic Instability; D006801:Humans; D007621:Karyotyping