Clinic for Gynecology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany.
Department of Otolaryngology, Head and Neck Surgery, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Berlin, Germany.
Biomed Pharmacother. 2019 Oct;118:109371. doi: 10.1016/j.biopha.2019.109371. Epub 2019 Aug 30.
Disulfiram (DSF) is a drug used for treatment of alcoholism that has also displayed promising anti-cancer activity. It unfolds its effects by inhibiting the enzyme activity of aldehyde dehydrogenase (ALDH) isoforms.
MTT assay, spheroid formation, clonogenicity assay, qRT-PCR, and ALDH enzyme activity analysis were performed using ovarian cancer cell lines IGROV1, SKOV3 and SKOV3IP1. Cell cycle analyses and measurement of intracellular reactive oxygen species (ROS) were carried out by flow cytometry. ALDH+ and ALDH- cells were isolated by FACS sorting.
ALDH activity was inhibited in ovarian cancer stem cells (the proportion of ALDH+ cells was reduced from 21.7% to 0.391%, 8.4% to 0%, 6.88% to 0.05% in cell lines IGROV1, SKOV3, and SKOV3IP1, respectively). DSF with or without the cofactor copper (Cu) exhibited cytotoxicity dose- and time-dependent and enhanced cisplatin-induced apoptosis. DSF + Cu increased intracellular ROS levels triggering apoptosis of ovarian cancer stem cells (CSC). Significantly more colony and spheroid formation was observed in ALDH+ compared with ALDH- cells (P < 0.01). Moreover, ALDH+ cells were more resistant to cisplatin treatment compared with ALDH-cells (P < 0.05) and also exhibited a lower basal level of ROS. However, no significant difference in ROS accumulation nor in cellular viability was observed in ALDH + cells in comparison to ALDH- cells after pre-treatment with DSF (0.08 μM).
Our findings provide evidence that DSF might be employed as a novel adjuvant chemotherapeutic agent in combination with cisplatin for treatment of ovarian cancer.
双硫仑（DSF）是一种用于治疗酗酒的药物，也显示出有前景的抗癌活性。它通过抑制醛脱氢酶（ALDH）同工酶的酶活性来发挥作用。
使用卵巢癌细胞系 IGROV1、SKOV3 和 SKOV3IP1 进行 MTT 测定、球体形成、集落形成能力测定、qRT-PCR 和 ALDH 酶活性分析。通过流式细胞术进行细胞周期分析和细胞内活性氧（ROS）的测量。通过 FACS 分选分离 ALDH+和 ALDH-细胞。
卵巢癌细胞中的 ALDH 活性受到抑制（细胞系 IGROV1、SKOV3 和 SKOV3IP1 中 ALDH+细胞的比例分别从 21.7%减少到 0.391%、8.4%减少到 0%、6.88%减少到 0.05%）。DSF 联合或不联合辅因子铜（Cu）表现出剂量和时间依赖性的细胞毒性，并增强顺铂诱导的细胞凋亡。DSF+Cu 增加细胞内 ROS 水平，触发卵巢癌细胞的凋亡（CSC）。与 ALDH-细胞相比，ALDH+细胞中观察到更多的集落和球体形成（P<0.01）。此外，与 ALDH-细胞相比，ALDH+细胞对顺铂治疗的耐药性更强（P<0.05），并且 ROS 水平也较低。然而，在使用 DSF（0.08μM）预处理后，与 ALDH-细胞相比，ALDH+细胞中 ROS 积累或细胞活力均未观察到显著差异。
我们的研究结果表明，DSF 可能被用作新型辅助化疗药物与顺铂联合用于治疗卵巢癌。
