The effects of nuclear factor kappa B decoy oligodeoxynucleotides on ciglitazone-induced differentiation of lung cancer cells A549
XU Yong-jia
Abstract
XU Yong-jia
Abstract
ObjectiveTo investigate the role of nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotides (ODNs) in the process of differential regulation of the lung cancer cells A549 by ciglitazone. MethodsNF-κB decoy ODNs were transfected into A549 cells with Lipofect AMINE 2000,and electrophoretic mobility shift assay(EMSA) was performed to investigate the activation of NF-κB, while the level of mdr1 was observed by Western blot. When ciglitazone was added, the proliferation was observed by growth curve and the differentiation of cells was observed by flow cytometry, and the expression of cyclinD1 was observed by Western blot. ResultsEMSA showed that the decoy ONDS-decreased the activation of NF-κB in A549, and Western blot showed that the decoy ONDS decreased the l evel of mdr1. Ciglitazone significantly inhibited the growth and induced the differentiation of A549 cells that were transfected with NF-κB decoy ODNs. There we re more cells arrested in G 1/G 0 phase and the expression of cyclinD1 was markedly down-regulated as compared to the control cells. ConclusionNF-κB decoy ODNs could enhance the effects of ciglitazone on proliferative suppression and differential induction of A549 cells.
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ObjectiveTo investigate the role of nuclear factor kappa B (NF-κB) decoy oligodeoxynucleotides (ODNs) in the process of differential regulation of the lung cancer cells A549 by ciglitazone. MethodsNF-κB decoy ODNs were transfected into A549 cells with Lipofect AMINE 2000,and electrophoretic mobility shift assay(EMSA) was performed to investigate the activation of NF-κB, while the level of mdr1 was observed by Western blot. When ciglitazone was added, the proliferation was observed by growth curve and the differentiation of cells was observed by flow cytometry, and the expression of cyclinD1 was observed by Western blot. ResultsEMSA showed that the decoy ONDS-decreased the activation of NF-κB in A549, and Western blot showed that the decoy ONDS decreased the l evel of mdr1. Ciglitazone significantly inhibited the growth and induced the differentiation of A549 cells that were transfected with NF-κB decoy ODNs. There we re more cells arrested in G 1/G 0 phase and the expression of cyclinD1 was markedly down-regulated as compared to the control cells. ConclusionNF-κB decoy ODNs could enhance the effects of ciglitazone on proliferative suppression and differential induction of A549 cells.
Key concepts: Decoy, A549 cell, Ciglitazone, Western blot, Transfection, Molecular biology, Flow cytometry, Medicine