[HPV16 E6E7 fragments transform immortalized human bronchial epithelial cells into neoplastic cells].
OBJECTIVE
To study the effects of HPV16 E6E7 fragments on the biological behavior of immortalized human bronchial epithelial cells TR.


METHODS
E6E7 fragments were constructed into retrovirus expression vector and then transfected into TR cells to observe changes in growth character and tumorigenesis in nude mice. IP-Western blots were done to analyze function of p27 protein as well as expression and phosphorylation of FAK and paxillin.


RESULTS
Southern, Northern, Western blot confirmed the stable expression of E6E7 fragments in puromycin-resistant cell strain TR/E6E7. TR/E6E7 proliferated faster, showed stronger growth ability in soft agar and eventually formed tumor after being inoculated into nude mice. IP-Western blot also showed sequestering of p27 protein from cyclin E/CDK2 complex, which implied increased activity of cyclin E/CDK2 and promoting G1 to S cell cycle.


CONCLUSIONS
TR/E6E7 cells formed tumor in nude mice indicating that HPV infection played a role in pulmonary carcinogenesis. Sequestering of p27 protein from cyclin E/CDK2 complex may be responsible for increased proliferation in cells transfected by HPV16 E6E7.
Zhang
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L
Department of Pathology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100730, China.
Liu
T
T
Liu
H
H
Gu
C
C
D000818:Animals; D015153:Blotting, Western; D001980:Bronchi / Q000166:cytology; D042846:CDC2-CDC28 Kinases; D018797:Cell Cycle Proteins / Q000378:metabolism; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D019927:Cyclin E / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2; D050760:Cyclin-Dependent Kinase Inhibitor p27; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000502:physiology; D050725:Papillomavirus E7 Proteins; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D012097:Repressor Proteins; D012190:Retroviridae / Q000235:genetics; D014162:Transfection; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000378:metabolism
C493961:Cdkn1b protein, mouse; D018797:Cell Cycle Proteins; D019927:Cyclin E; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D050760:Cyclin-Dependent Kinase Inhibitor p27; D017346:Protein Serine-Threonine Kinases; D042846:CDC2-CDC28 Kinases; C495894:CDK2 protein, human; C495893:Cdk2 protein, mouse; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases