Department of Molecular Biology of Cancer, Medical University of Lodz, Lodz, Poland.
Melanoma Res. 2010 Feb;20(1):21-34. doi: 10.1097/CMR.0b013e328333bbe4.
Metastatic melanoma is a highly life-threatening disease. The lack of response to radiotherapy and chemotherapy highlights the critical need for novel treatments. Parthenolide, an active component of feverfew (Tanacetum parthenium), inhibits proliferation and kills various cancer cells mainly by inducing apoptosis. The aim of the study was to examine anticancer effects of parthenolide in melanoma cells in vitro. The cytotoxicity of parthenolide was tested in melanoma cell lines and melanocytes, as well as melanoma cells directly derived from a surgical excision. Adherent cell proliferation was measured by tetrazolium derivative reduction assay. Loss of the plasma membrane integrity, hypodiploid events, reactive oxygen species generation, mitochondrial membrane potential dissipation, and caspase-3 activity were assessed by flow cytometric analysis. Microscopy was used to observe morphological changes and cell detachment. Parthenolide reduced the number of viable adherent cells in melanoma cultures. Half maximal inhibitory concentration values around 4 mumol/l were determined. Cell death accompanied by mitochondrial membrane depolarization and caspase-3 activation was observed as the result of parthenolide application. Interestingly, the melanoma cells from vertical growth phase and melanocytes were less susceptible to parthenolide-induced cell death than metastatic cells when drug concentration was at least 6 mumol/l. Reactive oxygen species level was not significantly increased in melanoma cells. However, preincubation of parthenolide with the thiol nucleophile N-acetyl-cysteine protected melanoma cells from parthenolide-induced cell death suggesting the reaction with intracellular thiols as the mechanism responsible for parthenolide activity. In conclusion, the observed anticancer activity makes parthenolide an attractive drug candidate for further testing in melanoma therapy.
转移性黑色素瘤是一种高度危及生命的疾病。对放疗和化疗缺乏反应突出表明需要新型治疗方法。小白菊内酯是小白菊(Tanacetum parthenium)的一种活性成分,通过诱导细胞凋亡,主要抑制增殖并杀死各种癌细胞。本研究旨在体外研究小白菊内酯对黑色素瘤细胞的抗癌作用。通过四唑盐还原试验检测小白菊内酯在黑色素瘤细胞系和黑素细胞以及直接从手术切除获得的黑色素瘤细胞中的细胞毒性。通过流式细胞术分析评估质膜完整性丧失、亚二倍体事件、活性氧物种生成、线粒体膜电位耗散和 caspase-3 活性。通过显微镜观察形态变化和细胞脱落。小白菊内酯减少了黑色素瘤培养物中贴壁细胞的数量。确定了约 4umol/L 的半最大抑制浓度值。应用小白菊内酯后观察到细胞死亡伴随着线粒体膜去极化和 caspase-3 激活。有趣的是,当药物浓度至少为 6umol/L 时,来自垂直生长阶段的黑色素瘤细胞和黑素细胞比转移性细胞对小白菊内酯诱导的细胞死亡的敏感性较低。黑色素瘤细胞中活性氧水平没有显著增加。然而,用硫醇亲核试剂 N-乙酰半胱氨酸预先孵育小白菊内酯可保护黑色素瘤细胞免受小白菊内酯诱导的细胞死亡,这表明与细胞内硫醇的反应是小白菊内酯活性的机制。总之,观察到的抗癌活性使小白菊内酯成为进一步在黑色素瘤治疗中进行测试的有吸引力的药物候选物。