[Effects of adenovirus-mediated p16 and p53 genes transfer on apoptosis and cell cycle of lung carcinoma cells].
OBJECTIVE
To explore the synergistic inhibition effect and apoptosis induction of p16 and p53 genes on lung carcinoma cells.


METHODS
E1-deficient and replication-defective recombinant p16 and p53 adenoviruses were generated by liposome-mediated co-transfection of recombinant plasmid pAdCMV-p16 or pAdCMV-p53 along with pJM17 and homologous recombination in 293 packaging cell. The lung cancer cell line H358, which had a homozygous deletion of p53 gene and no expression of p16 mRNA and protein, was infected with recombinant p16 and p53 adenovirus either individually or together.


RESULTS
Immunohistochemical analysis showed that recombinant adenovirus could transfer p53 gene into tumor cell with 98% efficiency. Western blot indicated that p16 and p53 proteins were expressed at a high level in infected H358 cell. Inhibition effect of p53 gene on proliferation of H358 cell was weaker than that of p16 gene, and the combined use of both genes could completely prevent the proliferation of H358 cell. In situ end-labeling and flow cytometry indicated that p16 could result in G(1) arrest of cell cycle and did not induce H358 cells to undergo apoptosis; p53 also induced apoptosis of few cells besides G(1) arrest; and the simultaneous use of p16 and p53 genes could induce marked apoptosis of H358 cells.


CONCLUSIONS
p16 and p53 genes possess the synergistic inhibiting effect on growth of lung cancer cells and can cooperate to induce apoptosis of H358 cell. The combined application of recombinant p16 and p53 adenoviruses can be used as a new strategy for cancer gene therapy.
Bai
X
X
Baixy@usa.net
Che
F
F
Li
J
J
Ma
Y
Y
Zhou
Y
Y
Zhai
J
J
Meng
L
L
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics* / Q000502:physiology; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000473:pathology / Q000628:therapy; D014162:Transfection; D014183:Transplantation, Heterologous; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000502:physiology
D019941:Cyclin-Dependent Kinase Inhibitor p16; D016159:Tumor Suppressor Protein p53