title: Chromosomal instability and APC gene mutations in human sporadic colorectal adenomas.
abstract: The mechanisms by which adenomatous polyposis coli (APC) gene mutations contribute to colorectal tumourigenesis and progression are still not fully understood. Using in vitro mouse embryonic stem cells, APC mutations have been proposed to dysregulate the interactions between kinetochores and microtubules during mitosis, leading to chromosomal instability (CIN) and aneuploidy. A link between APC mutations and aneuploidy in vivo among human sporadic colorectal adenomas has not been reported previously and was therefore investigated in the present series of 61 adenomas. Multi-parameter flow cytometry, based on scattering and fluorescence from the DNA-specific 4,6-diamidino-2-phenylindole-2-hydrochloride (DAPI) dye, which separates epithelial from stromal lymphocyte nuclei, was used to evaluate the DNA index (DI) and to sort epithelial nuclei. Additionally, DNA extracted from these sorted nuclei was used to analyse APC mutations by DNA sequencing. Aneuploidy was present in 20 of 61 adenomas (33%), with 15 of these 20 cases (75%) having a near-diploid DI (DI different from 1 and less than 1.3). APC mutations were detected in 19 adenomas (31%): 12 were within or downstream of the mutation cluster region (MCR), roughly defined by codons 1200-1500, and seven were upstream of the MCR. Overall, the prevalence of aneuploidy in APC wild-type and mutated adenomas was 26% and 47%, respectively, and no statistically significant association was found between APC status and DI (p = 0.142). However, when APC mutations were subdivided into two groups, ie occurring within/downstream of the MCR and upstream of the MCR, the association of APC mutations within and downstream of the MCR with aneuploidy was statistically significant (p = 0.017). In conclusion, the present data suggest that the type of APC mutation may play a role in the origin of CIN in vivo in human sporadic colorectal adenomas and that APC mutations within and downstream of the MCR, and large-scale chromosomal alterations, may co-operate in the progression of a subgroup of adenomas.
authors.affiliation: Department of Oncogenesis, National Institute for Cancer Research, Genoa, Italy. walter.giaretti@istge.it
authors.forename: Walter
authors.initials: W
authors.lastname: Giaretti
authors.forename: Tiziana
authors.initials: T
authors.lastname: Venesio
authors.forename: Claudia
authors.initials: C
authors.lastname: Prevosto
authors.forename: Federica
authors.initials: F
authors.lastname: Lombardo
authors.forename: Jenny
authors.initials: J
authors.lastname: Ceccarelli
authors.forename: Silvia
authors.initials: S
authors.lastname: Molinu
authors.forename: Mauro
authors.initials: M
authors.lastname: Risio
mesh_terms: D000236:Adenoma / Q000235:genetics*; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000782:Aneuploidy; D002467:Cell Nucleus / Q000502:physiology; D043171:Chromosomal Instability / Q000235:genetics*; D003062:Codon / Q000235:genetics; D015179:Colorectal Neoplasms / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics; D004847:Epithelial Cells / Q000502:physiology; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D017491:Genes, APC / Q000502:physiology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics
chemical_list: D003062:Codon; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins