University of Miami, Sylvester Comprehensive Cancer Center, Miami, FL, USA; Medical Oncology Department, Saint-Antoine Hospital, Public Assistance of Paris Hospitals, Pierre and Marie Curie University, Paris VI, Paris, France.
University of Texas M. D. Anderson Cancer Center, Houston, TX, USA.
Mol Oncol. 2014 Dec;8(8):1458-68. doi: 10.1016/j.molonc.2014.05.016. Epub 2014 Jun 10.
A paradigm shift has occurred in the last decade from chemotherapy to targeted therapy for the management of many patients with advanced sarcoma. This work identifies a combination of targeted agents and doxorubicin that are effective against small cell sarcoma cell lines. Three small cell sarcoma cell lines were studied: RD18 (rhabdomyosarcoma), A204 (undifferentiated sarcoma) and TC 71 (Ewing's sarcoma). Each cell line was exposed to increasing concentrations of vorinostat (HDAC inhibitor), 17-DMAG (HSP90 inhibitor), abacavir (anti-telomerase) or sorafenib (tyrosine kinase inhibitor) alone, combined with one another, or combined with doxorubicin. Cell viability, cell cycle analysis and apoptosis were assessed by MTS assay, propidium iodide-Annexin V staining, and caspase 3/7 activity, respectively. The Chou and Talalay combination index (CI) was used to determine whether the effects were additive (CI = 1), synergistic (CI < 1) or antagonistic (CI > 1). In monotherapy, targeted agents achieved 30-90% reductions in viability, with the exception of abacavir. Dual-targeted combination therapies with vorinostat, sorafenib and 17-DMAG demonstrated synergy. Abacavir was antagonistic with every other drug and was not further studied. Both vorinostat and 17-DMAG synergized with doxorubicin, achieving 60% cell killing compared to 12% with doxorubicin alone. No synergy was observed for sorafenib with doxorubicin. The triple therapy vorinostat, 17-DMAG and doxorubicin did not show synergy, but increased the subG1 population at 24H, from 30% to 70% compared to monotherapies with an increase in apoptosis. This work provides evidence of synergy of combinations of vorinostat, 17-DMAG and sorafenib in small cell sarcoma. In addition to doxorubicin, these combinations enhance doxorubicin cytotoxicity at therapeutically relevant concentrations.
在过去的十年中，针对许多晚期肉瘤患者的治疗，从化疗到靶向治疗已经发生了范式转变。这项工作确定了靶向药物和阿霉素联合使用对小细胞肉瘤细胞系有效。研究了三种小细胞肉瘤细胞系：RD18（横纹肌肉瘤）、A204（未分化肉瘤）和 TC 71（尤因肉瘤）。每种细胞系都暴露于越来越高浓度的伏立诺他（HDAC 抑制剂）、17-DMAG（HSP90 抑制剂）、阿巴卡韦（端粒酶抑制剂）或索拉非尼（酪氨酸激酶抑制剂）单独、联合使用，或与阿霉素联合使用。通过 MTS 测定法评估细胞活力、细胞周期分析和细胞凋亡，分别用碘化丙啶-Annexin V 染色和 caspase 3/7 活性进行评估。Chou 和 Talalay 组合指数（CI）用于确定效果是否是相加的（CI = 1）、协同的（CI < 1）还是拮抗的（CI > 1）。在单药治疗中，除阿巴卡韦外，靶向药物使细胞活力降低 30-90%。伏立诺他、索拉非尼和 17-DMAG 的双重靶向联合治疗显示出协同作用。阿巴卡韦与其他每种药物均拮抗，不再进一步研究。伏立诺他和 17-DMAG 与阿霉素协同作用，与单独使用阿霉素相比，细胞杀伤率达到 60%，而单独使用阿霉素则达到 12%。索拉非尼与阿霉素无协同作用。伏立诺他、17-DMAG 和阿霉素的三联疗法没有显示出协同作用，但与单药治疗相比，在 24 小时时增加了亚 G1 群体，从 30%增加到 70%，同时增加了细胞凋亡。这项工作提供了伏立诺他、17-DMAG 和索拉非尼联合治疗小细胞肉瘤协同作用的证据。除阿霉素外，这些联合治疗还在治疗相关浓度下增强了阿霉素的细胞毒性。
