Transformation and immortalization of human keratinocytes by SV40.
We have studied the appearance of transformed properties following infection of human epidermal keratinocytes by the oncogenic virus SV40. Shortly after infection, only a small fraction of the cells are positive for SV40 T antigen by immunofluorescence; this fraction progressively increases upon serial subcultivation concomitant with an increase in plating/colony-forming efficiency and growth rate. The capacity of the cells to differentiate progressively decreases, as indicated by cytochemical staining and cornified cell-envelope formation induced by suspension in methyl cellulose. The infected cells enter a period of growth crisis characterized by cytopathology and cell death as the level of T antigen synthesis reaches about 90 percent positive cells at about the tenth serial passage. Viable cells emerging from the crisis period are found to exhibit anchorage-independent growth, as indicated by the formation of viable colonies in semisolid media, but there is considerable variability in colony formation among clones isolated from anchorage-independent populations. The emergent population also manifests phenotypic instability in terms of the appearance of variants, which, in contrast to uninfected cells, expresses a well-defined actin cytoskeleton. The infected cells eventually become "immortalized," as evidenced by an indefinite lifespan, i.e., replication capability maintained well beyond the ordinary time of senescence for uninfected cells. We present these findings in the context of a stage-specific model of epithelial transformation in vitro.
D000199:Actins / Q000032:analysis; D002472:Cell Transformation, Viral; D003599:Cytoskeleton / Q000648:ultrastructure; D000078404:Epidermal Cells; D005455:Fluorescent Antibody Technique; D006801:Humans; D010641:Phenotype; D013539:Simian virus 40
D000199:Actins