中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (8): 570-573.

• 论著 • 上一篇    下一篇

A375细胞外信号调节激酶通路与核因子κB通路的交互作用初探

秦绪艳1,臧运书1,潘敏2,袁梦姝1,郁博2,吴惠惠1   

  1. 1. 青岛大学附属医院皮肤科
    2. 青岛大学医学院附属医院
  • 收稿日期:2013-11-18 修回日期:2014-05-21 发布日期:2014-08-01
  • 通讯作者: 潘敏 E-mail:pan730412@126.com
  • 基金资助:
    山东省自然科学基金;山东省科技发展计划项目

Cross-talk between nuclear factor-κB and extracellular signal-regulated kinase signaling pathways in A375 human melanoma cells

  • Received:2013-11-18 Revised:2014-05-21 Published:2014-08-01

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摘要： 【摘要】 目的 探讨黑素瘤细胞A375细胞外信号调节激酶（ERK）通路与核因子κB（NF-κB）信号转导通路的交互作用。 方法 将培养的A375细胞分为对照组、选择性ERK信号通路抑制剂U0126（10 μmol/L、5 μmol/L）处理组和选择性NF-κB通路抑制剂BMS-345541（10 μmol/L、5 μmol/L）处理组，分别提取蛋白质和mRNA，用Western 印迹、RT-PCR观察U0126抑制ERK通路后，NF-κB P65、p-IκBα蛋白及NF-κB P65 mRNA的表达，观察BMS-345541抑制NF-κB通路后ERK1/2 、p-ERK1/2蛋白及ERK1 mRNA的表达。 结果 应用10 μmol/L、5 μmol/L U0126抑制ERK通路后，胞核NF-κB P65蛋白表达（分别为0.60 ± 0.04、0.56 ± 0.06）均较对照组（1.54 ± 0.15）减少（P < 0.01），其表达量与药物浓度无显著相关性（P > 0.01）；p-IκBα蛋白的表达（0.90 ± 0.05、0.70 ± 0.02）均较对照组（0.61 ± 0.03）增加（P < 0.01），两药物浓度组之间的表达量差异有统计学意义（P < 0.01）；NF-κB P65 mRNA的表达（0.79 ± 0.05，0.75 ± 0.04）均较对照组（0.86 ± 0.05）减少（P < 0.01）。应用10 μmol/L BMS-345541抑制NF-κB通路后，ERK1/2、p-ERK1/2蛋白及ERK1 mRNA的表达（0.73 ± 0.07、0.75 ± 0.09、1.51 ± 0.02）较对照组减少（P < 0.01），BMS-345541浓度为5 μmol/L时，ERK1/2、p-ERK1/2蛋白和ERK1 mRNA的表达（0.94 ± 0.11、0.99 ± 0.04、1.62 ± 0.03）较对照组的差异无统计学意义（均P > 0.05）。 结论 NF-κB通路和ERK通路在黑素瘤A375细胞中存在交互作用。

关键词: 黑色素瘤, NF-κB, 细胞外信号调节MAP激酶类, 信号转导

Abstract: Qin Xuyan, Zang Yunshu, Pan Min, Yuan Mengshu, Yu Bo, Wu Huihui. Department of Dermatology, Affiliated Hospital of Qingdao University, Qingdao 266003, China Corresponding author: Pan Min, Email: pan730412@126.com 【Abstract】 Objective To investigate the cross-talk between extracellular signal-regulated kinase （ERK） and nuclear factor κB （NF-κB） signal transduction pathways in A375 human melanoma cells. Methods Cultured A375 cells were randomly divided into 5 groups: control group receiving no treatment, two U0126 （a selective inhibitor of the ERK signaling pathway） groups treated with U0126 of 10 and 5 μmol/L, and two BMS-345541 groups treated with BMS-345541 of 10 and 5 μmol/L. After 24-hour treatment, Western blot and reverse transcription PCR were performed to measure the protein expressions of NF-κB P65, phosphorylated IκBα （p-IκBα）, ERK1/2, as well as p-ERK1/2, and the mRNA expressions of NF-κB P65 and ERK1, respectively. One-way analysis of variance and least significant difference （LSD）-t test were carried out for statistical analysis. Results After 24 hours of treatment with U0126 of 10 and 5 μmol/L, a significant decrease was noted in the relative expression level of NF-κB p65 protein （0.60 ± 0.04 and 0.56 ± 0.06 vs. 1.54 ± 0.15, both P < 0.01） and mRNA （0.79 ± 0.05 and 0.75 ± 0.04 vs. 0.86 ± 0.05, both P < 0.01）, but a statistical increase in that of p-IκBα protein （0.90 ± 0.05 and 0.70 ± 0.02 vs. 0.61 ± 0.03, both P < 0.01） in the two U0126 groups compared with the control group; significant differences were observed in the expression level of p-IκBα protein （P < 0.01） but not in that of NF-κB p65 protein （P > 0.01） between the two U0126 groups. The relative expression levels of ERK1/2 and p-ERK1/2 proteins as well as ERK1 mRNA were significantly higher in the control A375 cells than those in the cells treated with BMS-345541 of 10 μmol/L （0.73 ± 0.07，0.75 ± 0.09，1.51 ± 0.02, all P < 0.01）, but similar to those treated with BMS-345541 of 5 μmol /L （0.94 ± 0.11, 0.99 ± 0.04，1.62 ± 0.03, all P > 0.05）. Conclusion There is a cross-talk between ERK and NF-κB signal transduction pathways in A375 melanoma cells.

Key words: Melanoma, NF-kappa B, Extracellular signal-regulated MAP kinase, Signal transducting

引用本文

秦绪艳 臧运书 潘敏 袁梦姝 郁博 吴惠惠. A375细胞外信号调节激酶通路与核因子κB通路的交互作用初探[J]. 中华皮肤科杂志, 2014,47(8):570-573. doi: