Zhang Hui-lan, Zhang Zhen-xiang, Xu Yong-jian
Respiratory Department of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zhonghua Nei Ke Za Zhi. 2004 Mar;43(3):201-4.
To investigate the role of nuclear factor kappa B (NF-kappaB) decoy oligodeoxynucleotides (ODNs) in the process of differential regulation of the lung cancer cells A549 by ciglitazone.
NF-kappaB 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-kappaB, 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.
EMSA showed that the decoy ONDS-decreased the activation of NF-kappaB in A549, and Western blot showed that the decoy ONDS decreased the level of mdr1. Ciglitazone significantly inhibited the growth and induced the differentiation of A549 cells that were transfected with NF-kappaB decoy ODNs. There were more cells arrested in G(1)/G(0) phase and the expression of cyclinD1 was markedly down-regulated as compared to the control cells.
NF-kappaB decoy ODNs could enhance the effects of ciglitazone on proliferative suppression and differential induction of A549 cells.
探讨核因子κB（NF-κB）诱饵寡脱氧核苷酸（ODNs）在噻唑烷二酮类药物对肺癌细胞A549的差异调节过程中的作用。
用脂质体2000将NF-κB诱饵ODNs转染至A549细胞，采用电泳迁移率变动分析（EMSA）检测NF-κB的激活情况，同时用蛋白质免疫印迹法观察mdr1的水平。加入噻唑烷二酮类药物后，通过生长曲线观察细胞增殖情况，用流式细胞术观察细胞分化情况，并用蛋白质免疫印迹法观察细胞周期蛋白D1（cyclinD1）的表达。
EMSA结果显示，诱饵ODNs降低了A549细胞中NF-κB的激活，蛋白质免疫印迹法显示诱饵ODNs降低了mdr1的水平。噻唑烷二酮类药物显著抑制了转染NF-κB诱饵ODNs的A549细胞的生长并诱导其分化。与对照细胞相比，更多细胞停滞于G(1)/G(0)期，且cyclinD1的表达明显下调。
NF-κB诱饵ODNs可增强噻唑烷二酮类药物对A549细胞增殖抑制和诱导分化的作用。
