[Roles of p53 in the interaction of p21 and cell cycle proteins induced by benzo [a] pyrene].

OBJECTIVE: To investigate the roles of p53 in the interaction of p21, cyclin D1 and CDK4 in human embryonic lung fibroblasts (HELFs) induced by benzo (a) pyrene. METHODS: p53-H group (cells transfected with p53 small interference RNA plasmid, p53 siRNA) and HELF/CMV group (cells transfected with CMV vector) were treated with 2 micromol/L B [a] P for 24 h, and HELF/CMV + PFT-alpha group (HELF/CMV cells were treated with p53 chemical inhibitor, Pifithrin-alpha) was treated with 2 micromol/L B [a] P and 20 micromol/L PFT-alpha for 24 h. The above three groups set up control groups, respectively. Western blot assay was used to check the levels of p53, phosphorylated p53 at 20 site (p53-ser20), p21, cyclin D1 and CDK4. Immunoprecipitation assay was used to investigate the roles of p53 in the interaction of p21, cyclin D1 and CDK4. RESULTS: After inhibition of p53 using PFT-alpha or siRNA, the high levels of p53, p53-ser 20 and p21 induced by B [a] P were markedly decreased. The change of cyclin D1 level was not obsevered and the level CDK4 was free of B [a] P. The combination of p21 and CDK 4 was increased after HELFs exposed to B [a] P, which can not be observed after inhibition p53. The combination of p21 and cyclin D1 was increased with or without the expression of p53 after HELFs exposed to B [a] P. The combination of cyclin D1 and CDK 4 was not affected by B [a] P. CONCLUSION: p53 can affect the combination of p21 and CDK4 in HELFs induced by B [a] P.