Abstract
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.
Conclusion:
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.
MeSH terms
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Animals
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Blotting, Western
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Bronchi / cytology
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CDC2-CDC28 Kinases*
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Cell Cycle Proteins / metabolism
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Cell Division / genetics
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Cell Division / physiology
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Cell Line
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Cell Transformation, Neoplastic / genetics*
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Cyclin E / metabolism
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Cyclin-Dependent Kinase 2
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Cyclin-Dependent Kinase Inhibitor p27
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Cyclin-Dependent Kinases / metabolism
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Epithelial Cells / metabolism*
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Female
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Genetic Vectors / genetics
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Humans
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Mice
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Mice, Inbred BALB C
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Mice, Nude
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Neoplasm Transplantation
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Neoplasms, Experimental / metabolism
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Neoplasms, Experimental / pathology
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Oncogene Proteins, Viral / genetics*
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Oncogene Proteins, Viral / physiology
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Papillomavirus E7 Proteins
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Protein Serine-Threonine Kinases / metabolism
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Repressor Proteins*
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Retroviridae / genetics
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Transfection
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Transplantation, Heterologous
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Tumor Cells, Cultured
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Tumor Suppressor Proteins / metabolism
Substances
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Cdkn1b protein, mouse
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Cell Cycle Proteins
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Cyclin E
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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 Proteins
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oncogene protein E7, Human papillomavirus type 16
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Cyclin-Dependent Kinase Inhibitor p27
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Protein Serine-Threonine Kinases
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CDC2-CDC28 Kinases
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CDK2 protein, human
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Cdk2 protein, mouse
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Cyclin-Dependent Kinase 2
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Cyclin-Dependent Kinases