SV40 transfection of human kidney epithelial cells and stability of chromosome 11.
Simian virus 40 (SV40) transformation has been used in a variety of mammalian cells and has been shown to extend their life span. We therefore decided to apply these results to normal kidney and tumoral cells derived from Wilms' patients. Wilms' tumour, a nephroblastoma which presents in early childhood, has been linked to deletions and rearrangements in chromosome 11. Analysis of gene structure in the 11p13 locus involved in the development of the tumour has been restricted by the very short life-span of the tumoral cells in vitro. We transfected normal kidney WT/NK, tumoral WT/T cells and human foetal kidney HFK cells with 2 SV40-derived plasmids SV3neo (pBR322-SV40-containing neomycin bacterial gene) and SVori- (pMK-origin mutated SV40). We isolated a high number of SV40-transfected cell lines. The efficiency of transfection appeared to be extremely low in WT/T cells compared with HFK and even WT/NK. The life span of the cell lines was increased in relation to their untransfected homologues. However, in all of the cell lines except 3, senescence occurred, after crisis step or not. We looked at different markers associated with SV40 transformation of mammalian cells and specifically with the presence of SV40 T antigen in the cells and its consequences: AIG, tumorigenicity, expression and insertion in genomic DNA of SV40 T antigen. Genetic studies involving karyotypic and restriction fragment length polymorphism (RFLP) analysis demonstrated that, despite a frequent pseudo-diploidy, the cell lines derived have conserved the 11p13 locus.
D000957:Antigens, Viral, Tumor / Q000032:analysis; D002460:Cell Line; D002472:Cell Transformation, Viral; D002869:Chromosome Aberrations; D002880:Chromosomes, Human, Pair 11; D006801:Humans; D007668:Kidney / Q000648:ultrastructure*; D007680:Kidney Neoplasms / Q000235:genetics; D012150:Polymorphism, Restriction Fragment Length; D011519:Proto-Oncogenes; D013539:Simian virus 40 / Q000235:genetics*; D014162:Transfection; D009396:Wilms Tumor / Q000235:genetics
D000957:Antigens, Viral, Tumor