TRANSFECTED C-HA-RAS ONCOGENE ENHANCES KARYOTYPIC INSTABILITY AND INTEGRATES PREDOMINANTLY IN ABERRANT CHROMOSOMES

A human colon tumor cell line, SW480, was transfected with the c-Ha-ras oncogene, the wild type c-Ha-ras gene, or the pSV2neo plasmid. Cytogenetic analysis and localization of chromosome integration sites were combined in an attempt to analyze the effects of transfection with the c-Ha-ras oncogene on the karyotype. All transfected cell lines showed new clonal chromosome abnormalities present in all cells, ranging from three new aberrations in pSV2neo-transfected SW480 cell lines to eight in c-Ha-ras oncogene-transfected SW480 cell lines. The level of expression of c-Ha-ras mRNA after transfection with the c-Ha-ras oncogene was positively correlated with increased genetic instability, reflected in enhanced karyotypic instability. A combination of banding and fluorescence in situ hybridization (FISH) was used to identify chromosome integration sites. Plasmids containing ras integrated predominantly in new structurally rearranged chromosomes (five of eight). Three of five integration sites in new structurally rearranged chromosomes were localized at or near translocation breakpoints situated in telomeric regions. Specific chromosomes were not involved in the chromosome rearrangements. The results indicate that 1) enhanced expression of c-Ha-ras mRNA correlates with an increase in genetic instability in c-Ha-ras oncogene-transfected SW480 cell lines, and 2) that no specific integration site was observed but ras-containing plasmids were located predominantly in aberrant chromosomes near or at translocation breakpoints involving telomeric bands.