ABSTRACT
Objective To investigate the effect of STAT3 decoy ODNs on the growth and apoptosis of hepatoma cells Hep G2 and the related mechanism.Methods STAT3 decoy ODNs,synthesized in vitro,was transferred into Hep G2 cells by cationic lipid.The cell growth curve was used to reflect the growth and proliferation capacity of Hep G2,and annexin-V/PI staining method was used to calculate apoptic cells by FCM.Real time-PCR was used to quantitatively detect the expression of bcl-xL,cyclin D1,and c-myc mRNA in Hep G2.Results FCM assay and fluorescence microscopy demonstrated that STAT3 decoy ODNs was successfully transferred into Hep G2 cells(90.2%).Decoy ODNs inhibted the growth of Hep G2 cells.After treated with decoy ODNs 48 h,the percentage of early apoptotic cells was(22.86 ? 2.63)% and that of late apoptotic cells was(23.00 ? 2.76)%.Real time-PCR indicated that Hep G2 cells transfected with STAT3 decoy ODNs had decreased the mRNA expression of bcl-xL,cyclin D1 and c-myc.Conclusions STAT3 decoy ODNs inhibited the growth of Hep G2 cells and induce the apoptosis.The mechanism may be associated with decreased expression of bcl-xL,cyclin D1 and c-myc mRNA.
ABSTRACT
Objective To study the inhibitory effects of circular dumbbell decoy oligodeoxynucleotides targeting NF-?B on TNF-? expression in cortical neurons under the oxygen glucose deprivation/reoxygenation(OGD/R).Methods The cultured neurons dissociated from the cortex of neonatal rats were observed on the 7th day after culture.To make the ischemia/reperfusion model in vitro,the neurons were exposed to OGD 4 h before cultured in normal culture media.The neurons were divided into 5 groups according to different processing methods 2 h before exposure to OGD,including normal group,OGD4 h/R6 h treated,NF-?B decoy ODNs treated,scrambled decoy ODNs treated and TransfastFastTM Transfection Reagent treated groups.The protein level of NF-?B P65 in primary neurons was detected by Western blotting.The protein and mRNA levels of TNF-? were detected by immunocytochemical method and RT-PCR,respectively.Results NF-?B decoy ODNs could effectively inhibit the expression levels of NF-?B P65 protein,TNF-? mRNA and protein in neurons(P