title: Immortalization of human esophageal keratinocytes by E6 and E7 of human papillomavirus type 16.
abstract: Transduction of human papillomavirus type 16 (HPV16) E6/E7 into primary culture of human esophageal keratinocytes using a recombinant adenovirus prolonged the life-span, while untreated cells senesced within 14-16 population doublings (PDLs). Up-regulation of telomerase activity and acquisition of serum-resistant growth were observed in the esophageal keratinocytes with extended life-span between 50 and 100 PDLs, and drastically increased after 100 PDLs. A keratinocyte sample with a polymorphism of Pro/Pro at codon 72 of p53 showed resistance to HPV16 E6/E7-induced life-span-extension and immortalization, in contrast to others with p53 polymorphisms of Arg/Arg or Arg/Pro, which did not. The high efficiency of E6/E7-induction by adenovirus vector also revealed the M1 and M2 stages of keratinocyte immortalization first described in this report.
authors.affiliation: Department of Molecular Virology, Chiba University, Inohana, School of Medicine, Chiba 260-8670, Japan. sasiyama@virolo3.m.chiba-u.ac.jp
authors.forename: H
authors.initials: H
authors.lastname: Sashiyama
authors.forename: Y
authors.initials: Y
authors.lastname: Shino
authors.forename: Y
authors.initials: Y
authors.lastname: Kawamata
authors.forename: Y
authors.initials: Y
authors.lastname: Tomita
authors.forename: N
authors.initials: N
authors.lastname: Ogawa
authors.forename: H
authors.initials: H
authors.lastname: Shimada
authors.forename: S
authors.initials: S
authors.lastname: Kobayashi
authors.forename: T
authors.initials: T
authors.lastname: Asano
authors.forename: T
authors.initials: T
authors.lastname: Ochiai
authors.forename: H
authors.initials: H
authors.lastname: Shirasawa
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D002118:Calcium / Q000494:pharmacology; D002471:Cell Transformation, Neoplastic / Q000378:metabolism* / Q000473:pathology; D002472:Cell Transformation, Viral; D003114:Colony-Forming Units Assay; D017931:DNA Primers / Q000737:chemistry; D004847:Epithelial Cells / Q000378:metabolism / Q000821:virology*; D004947:Esophagus / Q000378:metabolism / Q000821:virology*; D005822:Genetic Vectors; D006801:Humans; D017403:In Situ Hybridization; D066298:In Vitro Techniques; D007633:Keratins / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D027383:Papillomaviridae / Q000235:genetics*; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; D016615:Telomere / Q000378:metabolism; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D017931:DNA Primers; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; D016159:Tumor Suppressor Protein p53; C059731:oncogene protein E7, Human papillomavirus type 16; D007633:Keratins; D002118:Calcium