氧化槐定碱通过下调IKK/IκB/NF-κB信号通路抑制结直肠癌SW480细胞增殖

Oxysophoridine inhibits the proliferation of colorectal cancer SW480 cell by down-regulating IKK/IκB/NF-κB signaling pathway

目的:探讨氧化槐定碱(Oxysophoridine,OSR)抑制SW480细胞增殖的IKK/IκB/NF-κB信号通路机制。方法:采用MTT法分析2、4、8、16、32、64、128及256 mg/l浓度氧化槐定碱作用72 h对SW480细胞增殖的抑制作用,并计算半数抑制浓度(IC50);将25、50及100 mg/l氧化槐定碱作用于SW480细胞,绘制7 d生长曲线;观察细胞集落形成情况;采用RT-PCR和Western blot法检测细胞IKKβ、NF-κB、IκBαmRNA和蛋白表达水平。结果:MTT结果提示氧化槐定碱浓度为2~256 mg/l时,对SW480细胞72 h增殖抑制率为4.76%~81.90%,IC50=(79.04±5.30)mg/l;生长曲线提示氧化槐定碱25、50和100 mg/l对SW480细胞生长具有明显的抑制作用;显著抑制SW480细胞的集落形成;氧化槐定碱25、100 mg/l可下调SW480细胞NF-κB、IKKβmRNA和蛋白表达水平,上调IκBαmRNA和蛋白表达水平。结论:氧化槐定碱对SW480细胞生长、增殖具有抑制作用,其作用机制可能与抑制IKK/IκB/NF-κB信号通路有关。

Objective:To explore the proliferation inhibition effect of Oxysophoridine(OSR)on colorectal cancer SW480 cell by IKK/IκB/NF-κB signaling pathway.Methods:The proliferation inhibition effect of OSR(72 h)at the dose of 2,4,8,16,32,64,128 and 256 mg/l on the SW480 cells were analyzed by MTT assay and the half inhibitory concentration(IC50)was calculated.SW480 cells were treated with OSR(25,50 and 100 mg/l)for 7 days to draw the growth curve and to observe the colony formation.The expression levels of IKKβ、NF-κB、IκBαmRNA and protein were detected by RT-PCR and Western blot.Results:After OSR(2-256 mg/l)treated for 72 h,the inhibition rata of W480 cells proliferation was 4.76%-81.90%by MTT,and IC50 was 79.04±5.30 mg/l.Growth curves indicated that OSR(25,50 and 100 mg/l)had significant inhibitory effects on SW480 cells.Colony forming of SW480 cells was significantly inhibited.RT-PCR and Western blot results showed that OSR(25,50 mg/l)down-regulated the expression levels of IKKβ、NF-κB mRNA and protein in SW480 cells,and up-regulated the expression levels of IκBα.Conclusion:OSR can inhibit the growth and proliferation of SW480 cells,its mechanism may be related to down-regulating the IKK/IκB/NF-κB signaling pathway.