摘要
目的探讨穿心莲内酯对肿瘤细胞生长的作用及机制。方法人肺腺癌A549细胞分别与穿心莲内酯1.5～30μmol·L-1作用24h,以及穿心莲内酯30μmol·L-1作用4～24h。用MTT法检测A549细胞存活率;Western印迹法检测在肿瘤坏死因子α(TNF-α)10μg·L-1刺激下,穿心莲内酯30μmol·L-1对NF-κB信号通路中的相关蛋白NF-κB抑制因子α(IκBα)、磷酸化IκBα、IκB激酶β(IKKβ)和磷酸化IKKβ表达的影响;ELISA法检测穿心莲内酯对A549细胞核内NF-κBDNA结合活性的影响。结果穿心莲内酯的浓度和作用时间与A549细胞的存活率密切相关,穿心莲内酯30μmo·lL-1作用24h,A549细胞的存活率下降到(22.0±1.2)%,而穿心莲内酯1.5μmol·L-1作用24h或者穿心莲内酯30μmol·L-1作用4h对A549细胞的存活率几乎无影响。Western印迹法显示,穿心莲内酯能够抑制TNF-α诱导的NF-κB信号通路中IKKβ的磷酸化,抑制IκBα的磷酸化,推迟IκBα的降解,对IKKβ的表达无影响。穿心莲内酯还能够抑制TNF-α诱导的A549细胞核内NF-κBp65蛋白的DNA结合活性,抑制率达32%。结论穿心莲内酯通过影响NF-κB信号通路抑制A549细胞的生长。
OBJECTIVE To investigate the effect of andrographolide (Andro) on the nuclear factor-kappa B (NF-κB) signaling pathway in human lung adenocarcinoma cell line A549 and possible mechanisms. METHODS A549 cells were treated with Andro 1.5,3,6,15 and 30 μmol·L-1 for 24 h, or with Andro 30 μmol·L-1 for 4, 8, 12 and 24 h. MTT method was used to detect cell survival. In addition, A549 cells were divided into 2 groups: control group, in which cells were only stimulated with TNF-α 10 μg·L-1 for 0, 5, 15, and 30 min; Andro group, in which cells were pre-incubated with Andro 30 μmol·L-1 for 4 h before 30 minutes of stimulation of TNF-α 10 μg·L-1, to study the expression of NF-κB signaling proteins, including inhibitor of κB α (IκBα), phosphorylated IκBα, IκB kinase β(IKKβ) and phosphorylated IKKβ by Western blotting and NF-κB DNA binding activity by ELISA. RESULTS The concentration and duration of both Andro treatment were related to the survival rate of A549 cells that declined to (22.0±1.2)% after 24 h incubation with Andro 30 μmol·L-1. In addition, the treatment of Andro 1.5 μmol·L-1 for 24 h or Andro 30 μmol·L-1 for 4 h had no effect on the survival rate of A549 cells. Furthermore, Andro inhibited TNF-α-induced phosphorylation of IKKβ in A549 cells, reduced phosphorylation of IκBα, blocked the subsequent degradation of IκBα, and had no effect on the expression of IKKβ. Andro decreased the DNA binding activity of NF-κB p65 in the nucleus of A549 cells and the inhibitory rate was 32%. CONCLUSION Andro can inhibit the growth of A549 cells by blocking NF-κB signaling pathway.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2010年第2期106-110,共5页
Chinese Journal of Pharmacology and Toxicology
