Down-regulation of AIF protein level can reduce the apoptosis induced by TAp63γ / 肿瘤

ABSTRACT

Objective: To investigate the role of apoptosis-inducing factor (AIF) in TAp63γ-induced apoptosis. Methods: Plasmid pcDNA3. 1-TAp63γ was transfected into human esophageal squamous cancer EC9706 cells for 24 h. Apoptosis of EC9706 cells was detected by observing DNA fragmentation. The apoptotic rate was measured by flow cytometry. The translocation of AIF in EC9706 cells was studied with Mitochondrial/Cytosol/Nuclear extraction method. The plasmid that expressed hairpin RNA specific for AIF were constructed using chemosynthesis and gene recombination technology, and the interfering efficacy of AIF shRNA was detected by western blotting. After siRNA interference the apoptotic rate of EC9706 cells transfected with pcDNA3. 1-TAp63γ was analyzed by flow cytometry. Results: Agarose gel electrophoresis revealed that DNA ladder, a typical feature of apoptosis, was observed in the pcDNA3. 1-TAp63γ plasmid-transfected cells 24 later, but not in the pcDNA3. 1 plasmid-transfected cells. The apoptotic rate was 13.64% and 1. 37% after EC9707 cells were transfected with pcDNA3. 1-TAp63γ and pcDNA3. 1 plasmid, respectively. Signals of AIF-immunostaining were detected in the nucleus and cytosol proteins of pcDNA3. 1-TAp63γ transfection group, but not in pcDNA3. 1 transfection group. The plasmid expressing shRNA specific for AIF was successfully constructed and the interfering efficiency was about 80%. The apoptotic rate in AIF siRNA group was 4.52% at 24 h after transfection with pcDNA3. 1-TAp63γ. Conclusions: TAp63 γ gene expression induces the apoptosis of EC9706 cells. Down-regulation of AIF protein expression decreases the apoptosis induced by TAp63 γ.