Genetic instability of chromosome 3 in HPV-immortalized and tumorigenic human keratinocytes

The HPV-1811 cell line is derived from primary human foreskin keratinocytes that have been transfected with human papilloma virus type 18. At late passage, these cells produce invasive squamous cell carcinomas when injected into nude mice. A striking, but unstable, aberration of chromosome 3 occurs very early after establishment of the culture; a consistent rearrangement is observed concomitant with tumorigenicity. Using molecular cytogenetic techniques, we characterized the complex development of this aberration. A whole chromosome probe to this chromosome was made by linker-adapter PCR amplification of a single flow-sorted chromosome. Hybridization of this probe to normal metaphase chromosomes revealed the der (3) to be composed of chromosome 3, distal 13q, and 21q22. Hybridization of a 3q subtelomeric probe and a glycoprotein V probe which maps to 3qter indicated that this locus is duplicated in the final form of the chromosome, but that much instability occurs prior to its establishment. The ETS2 oncogene, which maps to 21q22, is translocated to the der(3) when the cell line becomes tumorigenic, but not prior to this time. Early-passage cells which have been induced to become tumorigenic by exposure to the carcinogen nitrosomethylurea also have the localization of the ETS2 at 3qter.