Disulfiram deregulates HIF-alpha subunits and blunts tumor adaptation to hypoxia in hepatoma cells

作者：Park Hye joon; Kim Min sung; Cho Kumsun; Yun Jang hyuk; Choi Yong joon; Cho Chung hyun^*

来源：Acta Pharmacologica Sinica, 2013, 34(9): 1208-1216.

DOI：10.1038/aps.2013.52

• disulfiram
• hepatoma
• hypoxia
• HIF-2
• VEGF
• angiogenesis

Aim: Disulfiram is an aldehyde dehydrogenase inhibitor that was used toto treat alcoholism and showed anticancer activity, activity, but its anticancer mechanism remains unclear. The aim of this study was toto investigate the effects of disulfiram on the hypoxia-inducible factor (HIF)-driven tumor adaptation toto hypoxia in vitro.
Methods: Hep3B, Huh7 and HepG2 hepatoma cells were incubated under normoxic (20% O-2) or hypoxic (1% O2) conditions for 16 h. The expression and activity of HIF-1 alpha and HIF-2 alpha proteins were evaluated using immunoblotting and luciferase reporter assay, assay, assay, respectively. Semi-quantitative RT-PCR was used toto analyze HIF-mediated gene expression. Endothelial tubule formation assay was used toto evaluate the anti-angiogenic effect.
Results: Hypoxia caused marked expression of HIF-1 alpha and HIF-1 alpha 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-2 alpha 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 mu mol/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-2 alpha in Hep3B cells reversed the inhibitory effects of disulfiram on hypoxia-inducedia gene expression and cell survival under hypoxia.
Conclusion: Disulfiram deregulates the HIF-mediated hypoxic signaling pathway in hepatoma cells, which may contribute toto its anticancer effect. Thus, disulfiram could be used toto treat solid tumors that grow in a HIF-dependent manner.

• 出版日期2013-9