The Effect and Mechanism of Celecoxib in Hypoxia-Induced Survivin Up-Regulation in HUVECs

Background/Aims: To investigate the roles of hypoxia-inducible factor 1 alpha (HIF-1 alpha), cyclooxygenase-2 (Cox-2) and its product, Prostaglandin E2 (PGE(2)), in the mechanisms underlying hypoxia-induced survivin expression in human umbilical vein endothelial cells (HUVECs) and to examine the effect of celecoxib, a selective Cox-2 inhibitor, on survivin expression. Methods: HUVECs were exposed to a normal (95% O-2) or hypoxic (3% O-2) environment for 24 hrs. We observed the localized expression of survivin, Cox-2 and HIF-1 alpha in HUVECs using immunocytochemistry and detected the inhibitory effects of celecoxib on the growth of HUVECs using an MTT assay. mRNA and protein levels of Cox-2, HIF-1 alpha and survivin were determined by real-time PCR and Western blot analysis under hypoxic conditions for 0, 6, 12, or 24 hrs. The time course changes of HIF-1 alpha and survivin protein expression induced by cobalt chloride (CoCl2) were studied using Western blot analysis. We then treated HUVECs under hypoxia for 24 hrs with celecoxib (a Cox-2 selective inhibitor), genistein (a HIF-1 alpha inhibitor) or exogenous PGE(2) to further investigate the changes in hypoxia-induced survivin expression. Results: Following 24 hrs of hypoxic treatment, cells exhibited strongly positive survivin, HIF-1 alpha and Cox-2 cytoplasmic staining. Celecoxib (65 mu M) effectively inhibited cell proliferation under hypoxic conditions. The protein and mRNA levels of Cox-2, HIF-1 alpha and survivin were increased under hypoxia. The patterns of HIF-1 alpha and survivin expression induced by CoCl2 were similar to those induced by exposure to hypoxia. Genistein partially blocked survivin expression. Celecoxib reversed the hypoxia-induced survivin expression, whereas the addition of PGE(2) partially restored this effect. Conclusions: Hypoxia-induced survivin expression in HUVECs may be mediated by dual interdependent mechanisms directly involving HIF-1 alpha and indirectly involving the Cox-2/PGE(2) pathways. Celecoxib may offset hypoxia-induced survivin expression. Copyright (C) 2015 S. Karger AG, Basel