Sehwa Girls' High School, 56-7 Sinbanpo-ro, Seocho-gu, Seoul 137-040, Korea.
Acta Pharmacol Sin. 2013 Sep;34(9):1208-16. doi: 10.1038/aps.2013.52. Epub 2013 Jul 15.
Disulfiram is an aldehyde dehydrogenase inhibitor that was used to treat alcoholism and showed anticancer activity, but its anticancer mechanism remains unclear. The aim of this study was to investigate the effects of disulfiram on the hypoxia-inducible factor (HIF)-driven tumor adaptation to hypoxia in vitro.
Hep3B, Huh7 and HepG2 hepatoma cells were incubated under normoxic (20% O2) or hypoxic (1% O2) conditions for 16 h. The expression and activity of HIF-1α and HIF-2α proteins were evaluated using immunoblotting and luciferase reporter assay, respectively. Semi-quantitative RT-PCR was used to analyze HIF-mediated gene expression. Endothelial tubule formation assay was used to evaluate the anti-angiogenic effect.
Hypoxia caused marked expression of HIF-1α and HIF-1α in the 3 hepatoma cell lines, dramatically increased HIF activity and induced the expression of HIF downstream genes (EPO, CA9, VEGF-A and PDK1) in Hep3B cells. HIF-2α expression was positively correlated with the induction of hypoxic genes (CA9, VEGF-A and PDK1). Moreover, hypoxia markedly increased VEGF production and angiogenic potential of Hep3B cells. Disulfiram (0.3 to 2 μmol/L) inhibited hypoxia-induced gene expression and HIF activity in a dose-dependent manner. Disulfiram more effectively suppressed the viability of Hep3B cells under hypoxia, but it did not affect the cell cycle. Overexpression of HIF-2α in Hep3B cells reversed the inhibitory effects of disulfiram on hypoxia-induced gene expression and cell survival under hypoxia.
Disulfiram deregulates the HIF-mediated hypoxic signaling pathway in hepatoma cells, which may contribute to its anticancer effect. Thus, disulfiram could be used to treat solid tumors that grow in a HIF-dependent manner.
双硫仑是一种醛脱氢酶抑制剂，曾用于治疗酒精中毒，并显示出抗癌活性，但抗癌机制尚不清楚。本研究旨在探讨双硫仑对体外缺氧诱导因子（HIF）驱动的肿瘤适应缺氧的影响。
将 Hep3B、Huh7 和 HepG2 肝癌细胞在常氧（20% O2）或缺氧（1% O2）条件下孵育 16 小时。使用免疫印迹和荧光素酶报告基因检测分别评估 HIF-1α 和 HIF-2α 蛋白的表达和活性。半定量 RT-PCR 用于分析 HIF 介导的基因表达。内皮小管形成试验用于评估抗血管生成作用。
缺氧导致 3 种肝癌细胞系中 HIF-1α 和 HIF-1α 的明显表达，显著增加 HIF 活性，并诱导 Hep3B 细胞中 HIF 下游基因（EPO、CA9、VEGF-A 和 PDK1）的表达。HIF-2α 的表达与缺氧基因（CA9、VEGF-A 和 PDK1）的诱导呈正相关。此外，缺氧显著增加了 Hep3B 细胞中 VEGF 的产生和血管生成潜能。双硫仑（0.3 至 2 μmol/L）以剂量依赖性方式抑制缺氧诱导的基因表达和 HIF 活性。双硫仑更有效地抑制缺氧条件下 Hep3B 细胞的活力，但不影响细胞周期。在 Hep3B 细胞中转染过量表达的 HIF-2α 可逆转双硫仑对缺氧诱导基因表达和细胞在缺氧条件下存活的抑制作用。
双硫仑调节肝癌细胞中 HIF 介导的缺氧信号通路，这可能有助于其抗癌作用。因此，双硫仑可用于治疗以 HIF 依赖方式生长的实体瘤。
