[Effect of mouse p53 minigene on lung cancer cells with different 172 structures regulated by tetracycline].
Yanyan Ma, Binquan Wu, Jieming Xie, You J, Jianjun Liu, Xu Cui, J Wang, P Hui
Abstract
Yanyan Ma, Binquan Wu, Jieming Xie, You J, Jianjun Liu, Xu Cui, J Wang, P Hui
Abstract
OBJECTIVE: To investigate the effect of mouse 172 wild-type p53 (Arg), pseudo-wild-mutant-type p53 (Arg-->Leu) and mutant-type p53 (Arg-->His) induced by absence of tetracycline on the growth of PG cell line. METHODS: Three variant types of p53 minigene were sub-cloned by gene recombination into an expression vector which was controlled by tetracycline. Through LipofectAMINE, the vectors were transfected into p53 defective PG (248CGG-->CTT) cells, and the transfectants were screened in the selecting medium containing puromycin. Tumor suppressing effects were studied by MTT absorption, flow cytometry and Western blotting. RESULTS: Wild-type p53 and pseudo-wild-mutant-type p53 could lead cells to decrease their growth rates, arrest cell cycle and transactivation of p21(WAF1). Mutant-type p53 was defective in tumor suppression. CONCLUSION: Wild-type p53 and pseudo-wild-type p53 may inhibit cell growth and induce cell cycle arrest. Some p53 variants such as 172 Arg-->Leu can still retain the tumor suppression function of the wild-type.
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OBJECTIVE: To investigate the effect of mouse 172 wild-type p53 (Arg), pseudo-wild-mutant-type p53 (Arg-->Leu) and mutant-type p53 (Arg-->His) induced by absence of tetracycline on the growth of PG cell line. METHODS: Three variant types of p53 minigene were sub-cloned by gene recombination into an expression vector which was controlled by tetracycline. Through LipofectAMINE, the vectors were transfected into p53 defective PG (248CGG-->CTT) cells, and the transfectants were screened in the selecting medium containing puromycin. Tumor suppressing effects were studied by MTT absorption, flow cytometry and Western blotting. RESULTS: Wild-type p53 and pseudo-wild-mutant-type p53 could lead cells to decrease their growth rates, arrest cell cycle and transactivation of p21(WAF1). Mutant-type p53 was defective in tumor suppression. CONCLUSION: Wild-type p53 and pseudo-wild-type p53 may inhibit cell growth and induce cell cycle arrest. Some p53 variants such as 172 Arg-->Leu can still retain the tumor suppression function of the wild-type.
Key concepts: Wild type, Transactivation, Transfection, Molecular biology, Minigene, Mutant, Biology, Lipofectamine