title: Intratumor heterogeneity of K-ras2 mutations in colorectal adenocarcinomas: association with degree of DNA aneuploidy.
abstract: Detailed information about intratumor K-ras2 mutations in colorectal adenocarcinomas and a possible association with DNA content heterogeneity is still lacking. DNA diploid and aneuploid subclones, detected among multiple histologically selected primary sectors (57 superficial and 40 deep) and 9 lymph node metastases, were flow cytometrically sorted and separately submitted to codons 12-13 K-ras2 mutation spectrum analysis. DNA aneuploidy was absent among 20 near and 20 distant mucosa sites and present in 7/9 lymph node metastases and in 17/19 primary tumors (90%). Primary intratumor DNA multiclonality was approximately 50%. Degree of DNA aneuploidy (DNA Index) distribution was nonrandom and showed peaks at approximate mean DNA Index values 1.2, 1.5, and 1.8. K-ras2 mutations were detected in 0/20 mucosa cases, in 2/9 lymph node metastases, and in 9/19 adenocarcinomas (47%). No more than one mutation type per tumor was detected. Intratumor distribution of K-ras2 mutations was homogeneous in 6 and heterogeneous in 3 cases. Homogeneous distribution was associated with DNA near-diploid aneuploidy. K-ras2 mutations were strongly associated with DNA Index in the near-diploid region (83%) and almost absent (5%) among DNA near-triploid subclones (P = 0.0001). K-ras2 mutation intratumor heterogeneity indicates that sampling of the tumor may be a critical step and suggests that K-ras2 activation may be a late event in a subgroup of tumors. Our data also suggest the existence of an early process of the colorectal carcinogenesis that favors both K-ras2 mutations and DNA near-diploid aneuploidy. Onset of DNA near-triploid subclones appears, instead, to be independent from K-ras2 activation.
authors.affiliation: Laboratory of Biophysics and Cytometry, National Institute for Cancer Research (IST), Genoa, Italy.
authors.forename: W
authors.initials: W
authors.lastname: Giaretti
authors.forename: R
authors.initials: R
authors.lastname: Monaco
authors.forename: N
authors.initials: N
authors.lastname: Pujic
authors.forename: A
authors.initials: A
authors.lastname: Rapallo
authors.forename: S
authors.initials: S
authors.lastname: Nigro
authors.forename: E
authors.initials: E
authors.lastname: Geido
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology / Q000556:secondary; D003001:Cloning, Molecular; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000473:pathology; D004273:DNA, Neoplasm / Q000302:isolation & purification*; D005434:Flow Cytometry; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D008207:Lymphatic Metastasis; D011003:Ploidies; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: D004273:DNA, Neoplasm