Modulatory properties of various natural chemopreventive agents on the activation of NF-κB signaling pathway
Modulatory properties of various natural chemopreventive agents on the activation of NF-κB signaling pathway
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DOI：
10.1023/b:pham.0000022413.43212.cf
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发表时间：
2004-04-01
影响因子：
3.7
通讯作者：
Kong, ANT
中科院分区：
文献类型：
--
作者：
Jeong, WS;Kim, IW;Kong, ANT
目的。研究并比较几种天然化学预防药物对人HT-29结肠癌细胞核因子κ B （nf - κ B）转录激活的影响。本研究检测了十字花科蔬菜中的天然化学预防化合物异硫氰酸酯（ITCs）、绿茶中的类黄酮、红酒中的白藜芦醇（RES）和原花青素二聚体，以及姜黄咖喱食品中的姜黄素（CUR）。用NF-kappaB荧光素酶构建物稳定转染HT-29细胞，选择稳定克隆。选择其中一个克隆HT-29 N9细胞，用不同浓度的天然化学预防剂处理，随后用NF-kappaB刺激脂多糖（LPS）刺激，并测量荧光素酶活性。Western blot分析天然化学预防剂处理后磷酸化的IkappaBalpha。通过非放射性细胞增殖MTS试验[3-（4,5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺苯基）- 2h -四氮唑，内盐]、台苯蓝染色和caspase试验评估这些药物对细胞活力和凋亡的影响。在nf - kappab -荧光素酶实验中，用天然化学预防化合物处理导致不同的反应。异硫氰酸苯乙酯（PEITC）、萝卜硫素（SUL）、异硫氰酸烯丙基（AITC）和姜黄素（CUR）等ITCs强烈抑制lps诱导的nf - kappab -荧光素酶的激活，而RES在低剂量下增加了激活，但在高剂量下抑制了激活，茶黄酮和原花青素二聚体对nf - kappab -荧光素酶的作用很小或没有作用。ITCs、CUR、(-)-表没食子儿茶素-3-没食子酸酯（EGCG）和RES降低了lps诱导的ikappabα磷酸化。此外，在MTS实验中，PEITC、SUL和CUR也能有效抑制细胞生长。化学预防化合物可诱导Caspase-3活性，但在48 h内，不同化合物对Caspase-3的激活动力学存在差异。这些结果表明，天然化学预防药物在结肠癌和/或结肠癌的信号转导途径上具有不同的生物学功能。
Purpose. To study and compare effects of selected natural chemopreventive agents on the transcription activation of nuclear factor-kappa B (NF-kappaB) in human HT-29 colon cancer cells.Methods. The natural chemopreventive compounds isothiocyanates (ITCs) found in cruciferous vegetables, flavonoids found in green tea, resveratrol ( RES) and procyanidin dimers found in red wine, and curcumin ( CUR) found in turmeric curry food were examined in this study. HT-29 cells were stably transfected with NF-kappaB luciferase construct, and stable clones were selected. One of the clones, HT-29 N9 cells, was selected and treated with various concentrations of the natural chemopreventive agents and subsequently challenged with NF-kappaB stimulator lipopolysaccharide (LPS), and the luciferase activities were measured. Western blot analysis of phosphorylated IkappaBalpha was performed after treatments with the natural chemopreventive agents. The effects of these agents on cell viability and apoptosis were also evaluated by a nonradioactive cell proliferation MTS assay [3-(4,5-dimethylthiazol-2-yl)-5-(3-arboxymethoxyphenyl)-2-(4-sulfophenyl)-2H- tetrazolium, inner salt], Trypan blue staining, and caspase assay.Results. Treatments with the natural chemopreventive compounds resulted in different responses in the NF-kappaB-luciferase assay. ITCs such as phenethyl isothiocyanate ( PEITC), sulforaphane (SUL), allyl isothiocyanate ( AITC), and curcumin ( CUR) strongly inhibited LPS-induced NF-kappaB-luciferase activations, whereas RES increased activation at lower dose, but inhibited activation at higher dose, and tea flavonoids and procyanidin dimers had little or no effects. ITCs, CUR, (-)- epigallocatechin-3-gallate ( EGCG), and RES reduced LPS-induced IkappaBalpha phosphorylation. Furthermore, in the MTS assay, PEITC, SUL, and CUR also potently inhibited cell growth. Caspase-3 activity was induced by chemopreventive compounds, however, the kinetics of caspase-3 activation varied between these compounds within the 48-h time period.Conclusions. These results suggest that natural chemopreventive agents have differential biological functions on the signal transduction pathways in the colon and/or colon cancer.