Related Experiment Videos
[HPV16 E6E7 fragments transform immortalized human bronchial epithelial cells into neoplastic cells]
1Department of Pathology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100730, China.
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.