Cell context-dependent activities of parthenolide in primary and metastatic melanoma cells
Cell context-dependent activities of parthenolide in primary and metastatic melanoma cells
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DOI：
10.1111/j.1476-5381.2010.00749.x
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发表时间：
2010-07-01
影响因子：
7.3
通讯作者：
Wozniak, M.
中科院分区：
文献类型：
--
作者：
Czyz, M.;Lesiak-Mieczkowska, K.;Wozniak, M.
背景和目的：越来越多的证据表明NF-kappa B是黑色素瘤转移和化疗耐药增加的重要因素。在这里，我们报告了parthenolide对未治疗、顺铂或TNF α治疗的黑色素瘤细胞系A375、1205Lu和WM793的影响，显示出不同水平的组成nf - κ B活性。实验方法：采用电泳迁移位移法评估NF-kappa B活性的变化，实时荧光定量PCR评估NF-kappa B调控基因的表达。流式细胞术检测细胞周期阻滞和凋亡情况。荧光显微镜观察细胞死亡情况。用划痕法测定迁移量，用Matrigel法测定侵袭性。关键结果：Parthenolide抑制黑色素瘤细胞的组成和诱导nf - κ B活性。这伴随着癌症相关基因的下调，这些基因的启动子中有NF-kappa b结合位点，包括：Bcl-X(L)、survivin、cyclin D1、interleukin 8和基质金属蛋白酶9。通过比较6 μ M孤子内酯的各种作用，发现1205Lu细胞最有效地诱导了与线粒体膜电位丧失相关的细胞凋亡，WM793细胞的细胞周期阻滞在G(0)/G(1)期，A375细胞的高转移电位明显降低。这些发现不仅反映了黑色素瘤细胞系之间nf - κ b调节基因基础表达的差异，还提示了其他parthenolide靶点参与细胞周期进展、迁移、侵袭和生存。结论：parthenolide对组成性和治疗性诱导的NF-kappa B通路的抑制可能对黑色素瘤的治疗有用，尽管在不同遗传背景的黑色素瘤细胞中诱导的变化的多样性表明该化合物具有上下文依赖的多药理学特性。
Background and purpose:Growing evidence implicates NF-kappa B as an important contributor to metastasis and increased chemoresistance of melanoma. Here, we report the effects of parthenolide on either untreated, cisplatin- or TNF alpha-treated melanoma cell lines A375, 1205Lu and WM793, exhibiting different levels of constitutive NF-kappa B activity.Experimental approach:Electrophoretic mobility shift assay was used to assess changes in NF-kappa B activity, and real-time PCR to evaluate expression of NF-kappa B-regulated genes. Cell cycle arrest and apoptosis were assessed by flow cytometry. Cell death was also visualized by fluorescence microscopy. Migration was determined by scratch assay and invasiveness by Matrigel assay.Key results:Parthenolide suppressed both constitutive and induced NF-kappa B activity in melanoma cells. This was accompanied by down-regulation of cancer-related genes, with NF-kappa B-binding sites in their promoters, including: Bcl-X(L) , survivin, cyclin D1, interleukin 8 and matrix metalloproteinase 9. When the various effects of 6 mu M parthenolide were compared, apoptosis associated with loss of mitochondrial membrane potential was most efficiently induced in 1205Lu cells, cell cycle arrest in G(0)/G(1) phase was observed in WM793 cells, and high metastatic potential was markedly reduced in A375 cells. These findings not only reflected differences between melanoma cell lines in basal expression of NF-kappa B-regulated genes, but also suggested other parthenolide targets involved in cell cycle progression, migration, invasiveness and survival.Conclusions:Inhibition of constitutive and therapeutically induced NF-kappa B pathway by parthenolide might be useful in the treatment of melanoma, although the diversity of changes induced in melanoma cells with different genetic backgrounds indicate context-dependent poly-pharmacological properties of this compound.