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中国癌症防治杂志 ›› 2017, Vol. 9 ›› Issue (1): 25-29.doi: 10.3969/j.issn.1674-5671.2017.01.06

• 麻醉镇痛与肿瘤专栏 • 上一篇    下一篇

布洛芬调控IKKβ/IκBα/NF-κB信号通路对肝癌细胞株增殖及凋亡的影响

  

  1. 广西医科大学附属肿瘤医院麻醉科
  • 出版日期:2017-02-25 发布日期:2017-03-24
  • 通讯作者: 潘灵辉。E-mail:plinghui@hotmail.com
  • 基金资助:
    广西科学研究与技术开发计划资助项目(2013BC26260)

Effect of ibuprofen on apoptosis and proliferation of hepatic cells via the IKKβ/IκBα/NF-κB signaling pathway

  1. Department of Anesthesiology,Affiliated Tumor Hospital of Guangxi Medical University
  • Online:2017-02-25 Published:2017-03-24
  • Contact: Pan Linghui.E-mail:plinghui@hotmail.com

摘要:

目的 探讨布洛芬调控IKKβ/IκBα/NF-κB信号通路对肝癌细胞增殖及凋亡的影响。方法 选择人肝癌QGY-7703细胞为研究对象,采用随机数字表法将QGY-7703细胞随机分为对照组(Qc组)和不同浓度布洛芬组,每组6例:Q1组(布洛芬作用浓度为250 μmol/L)、Q2组(布洛芬作用浓度为500 μmol/L)、Q3组(布洛芬作用浓度为1 000μmol/L)、Qc组加入等容量的RPMI-1640培养基。各组分别孵育24 h、48 h和72 h时,采用CCK-8法测定不同浓度及不同孵育时间细胞的增殖抑制情况,流式细胞术检测细胞凋亡情况;孵育48 h时,采用实时荧光定量PCR和Western blot检测各组细胞中IKKβ和Bcl-2基因mRNA和蛋白的变化以及磷酸化IKKβ(p-IKKβ)、磷酸化IκBα(p-IκBα)、NF-κBp65蛋白的表达情况。结果 与Qc组比较,Q1组、Q2组、Q3组细胞增殖抑制率及细胞凋亡率均增加,差异有统计学意义(P<0.05);Q1组、Q2组、Q3组细胞增殖抑制率及细胞凋亡率依照布洛芬作用浓度和时间的增加而依次增加,具有浓度和时间依赖性,差异有统计学意义(P<0.05);与Qc组比较,Q1组、Q2组、Q3组细胞IKKβ mRNA和Bcl-2 mRNA表达明显下调,差异有统计学意义(P<0.05);Q1组、Q2组、Q3组比较,细胞IKKβ mRNA和Bcl-2 mRNA表达依照布洛芬作用浓度的增加而依次下调,具有浓度依赖性,差异有统计学意义(P<0.05);与Qc组比较,Q1组、Q2组、Q3组细胞IKKβ、Bcl-2 、p-IKKβ、p-IκBα和NF-κBp65蛋白表达明显下调,差异有统计学意义(P<0.05);Q1组、Q2组、Q3组比较,细胞中IKKβ、Bcl-2 、p-IKKβ、p-IκBα和NF-κBp65蛋白的表达依照布洛芬作用浓度的增加而依次降低,具有浓度依赖性,差异有统计学意义(P<0.05)。结论 布洛芬可以抑制人肝癌QGY-7703细胞的增殖,促进细胞凋亡,可能与布洛芬调控细胞IKKβ/IκBα/NF-κB信号通路有关。

关键词: 肝肿瘤, 布洛芬, 肝癌QGY-7703细胞, 增殖, 凋亡, 信号通路

Abstract:

 Objective To investigate the effect of ibuprofen on apoptosis and proliferation of hepatic cells via the IKKβ/IκBα/NF-κB signaling pathway. Methods Human liver cancer QGY-7703 cells in logarithmic growth phase were seeded into culture plates or bottles. The cells were randomly divided into 4 groups using random number table(n=6 each):control group(Qc group) and 3 experimental groups given different ibuprofen concentrations (Q1,Q2,Q3 groups). The Q1,Q2 and Q3 groups were exposed to ibuprofen at respective final concentrations of 250,500 and 1000 μmol/L,while the control group was exposed to normovolemic RPM-l640 nutrient solution. Cell proliferation at 24,48 and 72 h later was measured using the CCK-8 assay,and apoptosis rate was measured using flow cytometry.After 48 h incubation,levels of IKKβ and Bcl-2 mRNA and protein,as well as levels of p-IKKβ, p-IκBα and NF-κBp65 protein,were measured using real-time PCR or Western blotting. Results Proliferation rates and apoptotic rates were significantly higher in the Q1,Q2 and Q3 groups than in the Qc group. These effects depended on ibuprofen dose and incubation time. Levels of IKKβ and Bcl-2 mRNA were significantly lower in Q1,Q2 and Q3 groups than in the Qc group,and this effect varied with ibuprofen concentration. Similarly,protein levels of IKKβ,Bcl-2,p-IKKβ,p-IκBα and NF-κBp65 were significantly lower in the Q1,Q2 and Q3 groups than in the Qc group. All these levels negatively correlated with ibuprofen concentration(P<0.05). Conclusions Ibuprofen can inhibit proliferation and induce apoptosis in human liver cancer QGY-7703 cells,and it may exert these effects by regulating IKKβ/IκBα/NF-κB signaling.

Key words: Liver neoplasms, Ibuprofen, Liver cancer QGY-7703 cells ,  Proliferation, Apoptosis , Signaling pathway