Sotillo Wendy Soria, Villagomez Rodrigo, Smiljanic Sandra, Huang Xiaoli, Malakpour Atena, Kempengren Sebastian, Rodrigo Gloria, Almanza Giovanna, Sterner Olov, Oredsson Stina
Department of Biology, Lund University, Lund, Sweden.
Molecular Biology and Biotechnology Institute, University Major of San Andrés, La Paz, Bolivia.
PLoS One. 2017 Sep 1;12(9):e0184304. doi: 10.1371/journal.pone.0184304. eCollection 2017.
New regimens are constantly being pursued in cancer treatment, especially in the context of treatment-resistant cancer stem cells (CSCs) that are assumed to be involved in cancer recurrence. Here, we investigated the anti-cancer activity of sesquiterpene lactones (SLs) isolated from Ambrosia arborescens and of synthetic derivatives in breast cancer cell lines, with a specific focus on activity against CSCs. The breast cancer cell lines MCF-7, JIMT-1, and HCC1937 and the normal-like breast epithelial cell line MCF-10A were treated with the SLs damsin and coronopilin, isolated from A. arborescens, and with ambrosin and dindol-01, synthesized using damsin. Inhibitory concentration 50 (IC50) values were obtained from dose-response curves. Based on IC50 values, doses in the μM range were used for investigating effects on cell proliferation, cell cycle phase distribution, cell death, micronuclei formation, and cell migration. Western blot analysis was used to investigate proteins involved in cell cycle regulation as well as in the NF-κB pathway since SLs have been shown to inhibit this transcription factor. Specific CSC effects were investigated using three CSC assays. All compounds inhibited cell proliferation; however, damsin and ambrosin were toxic at single-digit micromolar ranges, while higher concentrations were required for coronopilin and dindol-01. Of the four cell lines, the compounds had the least effect on the normal-like MCF-10A cells. The inhibition of cell proliferation can partly be explained by downregulation of cyclin-dependent kinase 2. All compounds inhibited tumour necrosis factor-α-induced translocation of NF-κB from the cytoplasm to the nucleus. Damsin and ambrosin treatment increased the number of micronuclei; moreover, another sign of DNA damage was the increased level of p53. Treatment with damsin and ambrosin decreased the CSC subpopulation and inhibited cell migration. Our results suggest that these compounds should be further investigated to find efficient CSC-inhibiting compounds.
癌症治疗中一直在探索新的治疗方案，尤其是在与癌症复发相关的耐药癌症干细胞（CSCs）方面。在此，我们研究了从豚草中分离出的倍半萜内酯（SLs）及其合成衍生物对乳腺癌细胞系的抗癌活性，特别关注其对CSCs的活性。用从豚草中分离出的SLs——damsin和coronopilin，以及用damsin合成的ambrosin和dindol - 01处理乳腺癌细胞系MCF - 7、JIMT - 1和HCC1937以及正常样乳腺上皮细胞系MCF - 10A。从剂量反应曲线获得半数抑制浓度（IC50）值。基于IC50值，使用微摩尔范围内的剂量来研究对细胞增殖、细胞周期阶段分布、细胞死亡、微核形成和细胞迁移的影响。由于SLs已被证明可抑制转录因子NF - κB，因此使用蛋白质印迹分析来研究参与细胞周期调控以及NF - κB途径的蛋白质。使用三种CSC检测方法研究特定的CSC效应。所有化合物均抑制细胞增殖；然而，damsin和ambrosin在个位数微摩尔范围内具有毒性，而coronopilin和dindol - 01则需要更高的浓度。在这四种细胞系中，这些化合物对正常样MCF - 10A细胞的影响最小。细胞增殖的抑制部分可通过细胞周期蛋白依赖性激酶2的下调来解释。所有化合物均抑制肿瘤坏死因子-α诱导的NF - κB从细胞质向细胞核的转位。Damsin和ambrosin处理增加了微核的数量；此外，DNA损伤的另一个迹象是p53水平的升高。用damsin和ambrosin处理可减少CSC亚群并抑制细胞迁移。我们的结果表明，应进一步研究这些化合物以找到有效的CSC抑制化合物。
