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.
MeSH terms
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Adenoviridae / genetics
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Calcium / pharmacology
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Cell Transformation, Neoplastic / metabolism*
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Cell Transformation, Neoplastic / pathology
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Cell Transformation, Viral
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Colony-Forming Units Assay
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DNA Primers / chemistry
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Epithelial Cells / metabolism
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Epithelial Cells / virology*
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Esophagus / metabolism
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Esophagus / virology*
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Genetic Vectors
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Humans
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In Situ Hybridization
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In Vitro Techniques
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Keratins / metabolism
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Microscopy, Phase-Contrast
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Oncogene Proteins, Viral / genetics*
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Papillomaviridae / genetics*
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Papillomavirus E7 Proteins
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Repressor Proteins*
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Telomere / metabolism
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Transfection
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Tumor Suppressor Protein p53 / genetics
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Tumor Suppressor Protein p53 / metabolism
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Up-Regulation
Substances
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DNA Primers
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E6 protein, Human papillomavirus type 16
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Oncogene Proteins, Viral
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Papillomavirus E7 Proteins
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Repressor Proteins
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Tumor Suppressor Protein p53
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oncogene protein E7, Human papillomavirus type 16
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Keratins
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Calcium