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p16 Gene Expression Mediated by Adenovirus and Its Effect on Cell Cycle Arrest
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Abstract:
Objective:To investigate the p16 gene expression in lung cancer cell line introduced by adenovirus, and to observe the effect of p16 gene on cell cycle arrest.Methods: The replication-deficient adenovirus Ad5-p16 was homologously recombined in 293 cells and used to infect the lung cancer cell line SPC-A1. The expression of P16 protein was identified by immunohistochemistry. Trypan blue staining was used to count the alive cells and to draw a cellular growth curve. The changes of cell cycle and apoptosis were analyzed by flow cytometry (FCM).Results:After 48 hours since the SPC-A1 cells were infected by the replication-deficient adenovirus Ad5-p16 at MOI=10, the positive rate of the cells for P16 expression was 71%. The cellular growth inhibition and cytopathic effects were present when the cells were infected with Ad5-p16 at MOI=100. The cell cycle arrest and the cell apoptosis were found by FCM.Conclusion:The expression of p16 gene in SPC-A1 lung cancer cells induced by the replication-deficient adenovirus may result in the inhibition of cell growth and the introduction of cell apoptosis. The strategy targeted to the tumor suppressor gene that regulates the cell cycle directly, obtained satisfactory results, and provided a reliable theory for lung cancer gene therapy.
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