ABSTRACT
Objective To investigate the role of Sp1 in the regulation of transcription of vascular endothelial growth factor ( VEGF ) in hypoxia-induced HepG2 cells.Methods Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effect of hypoxia on Sp1 protein and VEGF mRNA expression .Mithramycin A, the selective inhibitor of Sp1 and knock-out Sp1 gene with siRNA were used to examine the effect on VEGF mRNA expression induced by hypoxia in HepG2 cells.Results Hypoxia induced Sp1 protein and VEGF mRNA expression in HepG2 cells.Mithramycin A produced a concentration-dependent decrease of hypoxia-induced VEGF mRNA expression . After inhibition of Sp1 RNAi, VEGF mRNA expression was significantly repressed in HepG 2 cells.Conclusion Hypoxia can increase the expression of Sp1 protein and VEGF mRNA in HepG2 cells induced by hypoxia.The transcription of VEGF is regulated by Sp1 in hypoxia-induced HepG2 cells.
ABSTRACT
Objective To investigate the role of Sp1 in the regulation of transcription of vascular endothelial growth factor ( VEGF ) in hypoxia-induced HepG2 cells.Methods Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effect of hypoxia on Sp1 protein and VEGF mRNA expression .Mithramycin A, the selective inhibitor of Sp1 and knock-out Sp1 gene with siRNA were used to examine the effect on VEGF mRNA expression induced by hypoxia in HepG2 cells.Results Hypoxia induced Sp1 protein and VEGF mRNA expression in HepG2 cells.Mithramycin A produced a concentration-dependent decrease of hypoxia-induced VEGF mRNA expression . After inhibition of Sp1 RNAi, VEGF mRNA expression was significantly repressed in HepG 2 cells.Conclusion Hypoxia can increase the expression of Sp1 protein and VEGF mRNA in HepG2 cells induced by hypoxia.The transcription of VEGF is regulated by Sp1 in hypoxia-induced HepG2 cells.
ABSTRACT
AIM To investigate the protection of plicamycin on apoptosis in cerebellar granule neurons (CGN) of rat. METHODS TUNEL, Hoechst 33258 staining, agarose gel electrophoresis and fluorescein diacetate staining were used to detect morphological and biochemical characteristics of apoptosis in primary rat CGN. RESULTS Being pre-incubated with plicamycin for 1 h and lasting for 24 h, rat CGN apoptosis induced by low potassium basal modified Eagle′s medium for 24 h was inhibited in a plicamycin concentration-dependent manner. This effective concentrations of plicamycin were from 50 to 200 nmol·L-1, and the maximum inhibitory rate of plicamycin on CGN apoptosis was near 80% at 200 nmol·L-1. CONCLUSIONPlicamycin inhibits rat CGN apoptosis induced by low potassium.