Oncogene-induced replication stress preferentially targets common fragile sites in preneoplastic lesions. A genome-wide study.
Common fragile sites (CFSs) are regions of the genome prone to breakage by replication inhibitors (extrinsic replication stress). Recently, we and others observed that oncogene-induced replication stress (RS) induces DNA damage from the earliest stages of cancer. Our aim was to perform a genome-wide analysis in precancerous and cancerous experimental models to examine whether allelic imbalance occurs within CFSs. Subsequently, CFSs sequence characteristics were assessed. We used a growth-factor-induced human skin hyperplasia and a H-ras-induced mouse hyperplastic urothelium as preneoplastic models, along with an inducible U2OS-CDT1(Tet-ON) cancer cell line model, all bearing established oncogene-induced RS stimuli. Human DNA was analysed with Affymetrix SNP microarrays, while mouse DNA was analysed with Nimblegen array CGH. We studied 56 aphidicolin-type CFSs and 1914 regions of control, nonfragile DNA. Our theoretical in silico analysis spanned 2.16 billion nonoverlapping bases on human chromosomes 1-22. Our results provide direct experimental evidence indicating that genomic alterations were more common within CFSs in epidermal and urothelial preneoplastic lesions as well as in cancer. CFSs were on average less flexible than nonfragile regions, contained more guanine-cytosine (GC) and Alu sequences. Importantly, regions with loss-of-heterozygosity were also less flexible and had a higher Alu percentage.
Tsantoulis
P K
PK
Department of Histology and Embryology, Molecular Carcinogenesis Group, School of Medicine, University of Athens, Athens, Greece.
Kotsinas
A
A
Sfikakis
P P
PP
Evangelou
K
K
Sideridou
M
M
Levy
B
B
Mo
L
L
Kittas
C
C
Wu
X-R
XR
Papavassiliou
A G
AG
Gorgoulis
V G
VG
D000465:Algorithms; D000818:Animals; D045744:Cell Line, Tumor; D043283:Chromosome Fragile Sites; D004249:DNA Damage / Q000502:physiology; D004261:DNA Replication / Q000235:genetics; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D015894:Genome, Human; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis; D009857:Oncogenes / Q000502:physiology*; D011230:Precancerous Conditions / Q000235:genetics* / Q000473:pathology; D012878:Skin Neoplasms / Q000235:genetics; D014183:Transplantation, Heterologous