Normal human fibroblasts immortalized by introduction of human papillomavirus type 16 (HPV-16) E6-E7 genes.
This report demonstrates that normal human fibroblasts can be immortalized by the introduction of HPV-16 E6-E7 genes. We designed zinc-inducible expression plasmids with HPV-16 E6, E7 or both. Each plasmid was introduced into normal human fibroblasts (TIG-3 cells) using lipofection methods. Only transfectants with the HPV-16 E6-E7 zinc-inducible expression plasmid, which were cultured in medium supplemented with 100 microM ZnSO4, overcame crisis and could be cultured over 200 population doubling levels (PDLs). These cell lines showed the reactivation of telomerase after crisis, and morphological alterations were also observed.
Shiga
T
T
Department of Microbiology, School of Medicine, Chiba University, Chuo-ku, Japan.
Shirasawa
H
H
Shimizu
K
K
Dezawa
M
M
Masuda
Y
Y
Simizu
B
B
D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media; D005347:Fibroblasts / Q000166:cytology* / Q000821:virology*; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D050725:Papillomavirus E7 Proteins; D010957:Plasmids; D012097:Repressor Proteins; D019098:Telomerase / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured; D019287:Zinc Sulfate / Q000378:metabolism
D003470:Culture Media; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D019287:Zinc Sulfate; D019098:Telomerase