药学学报, 2017, 52(1): 51-57
引用本文:
黎玉华, 黄凌, 魏筱华, 温金华, 钟国平, 黄民, 毕惠嫦. PKC/NF-κB-PXR信号通路对P-糖蛋白基因表达的调控研究[J]. 药学学报, 2017, 52(1): 51-57.
LI Yu-hua, HUANG Ling, WEI Xiao-hua, WEN Jin-hua, ZHONG Guo-ping, HUANG Min, BI Hui-chang. Regulation of P-glycoprotein gene expression by PKC/NF-κB-PXR signaling pathway[J]. Acta Pharmaceutica Sinica, 2017, 52(1): 51-57.

PKC/NF-κB-PXR信号通路对P-糖蛋白基因表达的调控研究
黎玉华1,2, 黄凌3, 魏筱华1, 温金华1, 钟国平2, 黄民2, 毕惠嫦2
1. 南昌大学第一附属医院, 江西 南昌 330006;
2. 中山大学药学院, 广东 广州 510006;
3. 海南医学院药学院, 海南 海口 571199
摘要:
P-糖蛋白(P-gp)是ABC(ATP binding cassette)转运体家族重要成员,也是药物在机体内转运的重要载体。本文考察PKC/NF-κB-PXR信号途径对LS174T细胞中P-gp基因表达的调控作用。运用孕甾烷X受体(PXR)-MDR1双荧光报告基因实验探究PKC激动剂佛波酯(PMA)对LS174T细胞中MDR1荧光素酶活性的影响;分别采用real-time qPCR和Western blot检测PMA对LS174T细胞中P-gp mRNA表达、蛋白表达及NF-κB通路相关蛋白的影响。结果表明,PKC激动剂PMA能明显抑制PXR介导的P-gp荧光素酶活性、mRNA和蛋白表达,并能显著性增加胞内RelA/p65的核转位。此外,siRNA干扰实验结果显示,PKCα siRNA、RelA siRNA或PXR siRNA干扰均可显著削弱PMA对P-gp基因表达的下调作用。因此,PKC激动剂能显著抑制PXR介导的P-gp基因表达并伴随NF-κB激活,提示PKC/NF-κB-PXR信号通路对P-gp基因表达具有重要的调控作用。
关键词:    蛋白激酶C      核因子κB      孕甾烷X受体      P-糖蛋白      基因表达     
Regulation of P-glycoprotein gene expression by PKC/NF-κB-PXR signaling pathway
LI Yu-hua1,2, HUANG Ling3, WEI Xiao-hua1, WEN Jin-hua1, ZHONG Guo-ping2, HUANG Min2, BI Hui-chang2
1. The First Affiliated Hospital of Nanchang University, Nanchang 330006, China;
2. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China;
3. School of Pharmaceutical Sciences, Hainan Medical University, Haikou 571199, China
Abstract:
P-glycoprotein (P-gp), an ATP binding cassette protein, plays a major role in efflux transport of drugs and xenobiotics due to its abundant expression on several barriers. This study aimed to investigate the potential role of PKC/NF-κB-PXR signaling pathway in modulation of P-gp gene expression in human colon adenocarcinoma LS174T. The effect of PMA on MDR1 luciferase activity was investigated by PXR-MDR1 dual luciferase reporter gene assay. Real-time qPCR assay and Western blot analysis were used to study the gene expression of P-gp and NF-κB, respectively. Compared to the vehicle-treated group, PMA statistically decreased P-gp luciferase activity, mRNA expression and protein expression. Moreover, PMA treatment yielded a significant and dose-dependent increase in RelA/p65 translocation to nucleus. Meanwhile, a remarkable increase of the pho-IκBα status was observed in LS174T cells after treatment with PMA (1-100 nmol·L-1). In addition, knockdown of PKCα, NF-κB or PXR can significantly attenuate PMA-induced P-gp suppression.These results suggested that PKC/NF-κB-PXR signaling pathway might play crucial roles in modulation of P-gp gene expression.
Key words:    protein kinase C    nuclear factor κB    pregnane X receptor    p-glycoprotein    gene expression   
收稿日期: 2016-10-17
DOI: 10.16438/j.0513-4870.2016-1003
基金项目: 国家自然科学基金资助项目(81660618).
通讯作者: 毕惠嫦,Tel:86-20-39943470,Fax:86-20-39943000,E-mail:bihchang@mail.sysu.edu.cn
Email: bihchang@mail.sysu.edu.cn
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