Department of Thoracic Surgery, The Third Affiliated Hospital of Kunming Medical University, Kunming, China.
Cancer Center of Integrative Medicine, The Third Affiliated Hospital of Kunming Medical University, Kunming, China.
Biochem Biophys Res Commun. 2014 Apr 18;446(4):1010-6. doi: 10.1016/j.bbrc.2014.03.047. Epub 2014 Mar 19.
Non-small cell lung cancer (NSCLC) is the most common cause of cancer-related death in both men and women worldwide. Recently, Disulfiram has been reported to be able to inhibit glioblastoma, prostate, or breast cancer cell proliferation. In this study, the synergistic effect of Disulfiram and copper on NSCLC cell growth was investigated. Inhibition of cancer cell proliferation was detected by 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) assay and cell cycle analysis. Liquid colony formation and tumor spheroid formation assays were used to evaluate their effect on cancer cell clonogenicity. Real-time PCR was performed to test the mRNA level of cancer stem cell related genes. We found that Disulfiram or copper alone did not potently inhibit NSCLC cell proliferation in vitro. However, the presence of copper significantly enhanced inhibitory effect of Disulfiram on NSCLC cell growth, indicating a synergistic effect between Disulfiram and copper. Cell cycle analysis showed that Disulfiram/copper complex caused NSCLC cell cycle arrest in G2/M phase. Furthermore, Disulfiram/copper significantly increased the sensitivity of cisplatin in NSCLC cells tested by MTT assay. Liquid colony formation assay revealed that copper dramatically increased the inhibitory effect of Disulfiram on NSCLC cell colony forming ability. Disulfiram combined with copper significantly attenuated NSCLC cell spheroid formation and recuded the mRNA expression of lung cancer stem cell related genes. Our data suggest that Disulfiram/copper complex alone or combined with other chemotherapy is a potential therapeutic strategy for NSCLC patients.
非小细胞肺癌（NSCLC）是全球男性和女性癌症相关死亡的最常见原因。最近，报道发现双硫仑能够抑制神经胶质瘤、前列腺癌或乳腺癌细胞的增殖。在这项研究中，研究了双硫仑和铜对 NSCLC 细胞生长的协同作用。通过 1-(4,5-二甲基噻唑-2-基)-3,5-二苯基甲臜（MTT）测定和细胞周期分析检测癌细胞增殖的抑制作用。液体集落形成和肿瘤球体形成测定用于评估它们对癌细胞克隆形成能力的影响。实时 PCR 用于测试癌症干细胞相关基因的 mRNA 水平。我们发现，双硫仑或铜单独使用时不能有效地抑制 NSCLC 细胞的体外增殖。然而，铜的存在显著增强了双硫仑对 NSCLC 细胞生长的抑制作用，表明双硫仑和铜之间存在协同作用。细胞周期分析表明，双硫仑/铜复合物导致 NSCLC 细胞周期停滞在 G2/M 期。此外，双硫仑/铜复合物显著增加了 MTT 测定中 NSCLC 细胞对顺铂的敏感性。液体集落形成测定显示，铜显著增强了双硫仑对 NSCLC 细胞集落形成能力的抑制作用。双硫仑与铜联合显著减弱了 NSCLC 细胞球体的形成，并降低了肺癌干细胞相关基因的 mRNA 表达。我们的数据表明，双硫仑/铜复合物单独或与其他化疗联合是 NSCLC 患者的一种潜在治疗策略。
