Charité - Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Institute of Biochemistry, Charitéplatz 1, 10117, Berlin, Germany.
BMC Cancer. 2020 Jul 2;20(1):617. doi: 10.1186/s12885-020-07062-2.
Despite an improvement of prognosis in breast and colon cancer, the outcome of the metastatic disease is still severe. Microevolution of cancer cells often leads to drug resistance and tumor-recurrence. To target the driving forces of the tumor microevolution, we focused on synergistic drug combinations of selected compounds. The aim is to prevent the tumor from evolving in order to stabilize disease remission. To identify synergisms in a high number of compounds, we propose here a three-step concept that is cost efficient, independent of high-throughput machines and reliable in its predictions.
We created dose response curves using MTT- and SRB-assays with 14 different compounds in MCF-7, HT-29 and MDA-MB-231 cells. In order to efficiently screen for synergies, we developed a screening tool in which 14 drugs were combined (91 combinations) in MCF-7 and HT-29 using EC or less. The most promising combinations were verified by the method of Chou and Talalay.
All 14 compounds exhibit antitumor effects on each of the three cell lines. The screening tool resulted in 19 potential synergisms detected in HT-29 (20.9%) and 27 in MCF-7 (29.7%). Seven of the top combinations were further verified over the whole dose response curve, and for five combinations a significant synergy could be confirmed. The combination Nutlin-3 (inhibition of MDM2) and PX-478 (inhibition of HIF-1α) could be confirmed for all three cell lines. The same accounts for the combination of Dichloroacetate (PDH activation) and NHI-2 (LDH-A inhibition). Our screening method proved to be an efficient tool that is reliable in its projections.
The presented three-step concept proved to be cost- and time-efficient with respect to the resulting data. The newly found combinations show promising results in MCF-7, HT-29 and MDA-MB231 cancer cells.
尽管乳腺癌和结肠癌的预后有所改善，但转移性疾病的结局仍然很严重。癌细胞的微进化常常导致药物耐药和肿瘤复发。为了针对肿瘤微进化的驱动力，我们专注于选择化合物的协同药物组合。其目的是防止肿瘤进化，从而稳定疾病缓解。为了在大量化合物中识别协同作用，我们在这里提出了一个三步概念，该概念具有成本效益，不依赖于高通量机器，并且预测结果可靠。
我们使用 MTT 和 SRB 测定法在 MCF-7、HT-29 和 MDA-MB-231 细胞中用 14 种不同的化合物创建了剂量反应曲线。为了有效地筛选协同作用，我们开发了一种筛选工具，其中在 MCF-7 和 HT-29 中使用 EC 或更低的浓度组合了 14 种药物（91 种组合）。通过 Chou 和 Talalay 方法验证了最有前途的组合。
所有 14 种化合物在三种细胞系中均表现出抗肿瘤作用。筛选工具在 HT-29 中检测到 19 种潜在的协同作用（20.9％），在 MCF-7 中检测到 27 种（29.7％）。对前 7 种组合进行了整个剂量反应曲线的进一步验证，其中 5 种组合的协同作用得到了证实。在所有三种细胞系中都可以确认 Nutlin-3（MDM2 抑制）和 PX-478（HIF-1α 抑制）的组合。Dichloroacetate（PDH 激活）和 NHI-2（LDH-A 抑制）的组合也是如此。我们的筛选方法已被证明是一种高效可靠的工具。
所提出的三步概念在数据方面证明了其具有成本效益和时间效益。新发现的组合在 MCF-7、HT-29 和 MDA-MB231 癌细胞中显示出有希望的结果。
