Effects of silencing transcription factor AP-4 gene on apoptosis of endometrial cancer cells

BIE Yanhong; LI Na; WU Youming; WANG Jinsheng; GENG Wenjing; LAN Hailong; CHENG Guanghui

doi:10.3969/j.issn.1002-266X.2017.02.002

Abstract：Objective To investigate the effect of silencing transcription factor activating enhancer-binding protein 4 (TFAP-4)in vitro on apoptosis of endometrial cancer cells and its mechanism.Methods The human endometrial cancer cells (HEC-1-A and RL95-2 cells)were divided into NC group,si#1 group and si#2 group.The si#1 group and si#2 group were transfected with si-TFAP4 fragment 1 and si-TFAP4 fragment 2,respectively,and the NC group was transfected with control siRNA.The apoptosis was detected by Hoechest 33258 staining and the apoptosis rate by Annexin V-FITC/PI assay. The survivin (BIRC5)mRNA was detected by real-time PCR.Bioinformatics software was used to detect whether there were the AP-4 binding sites on the promoter of BIRC5.Chromatin immunoprecipitation was used to detect the binding between AP-4 and the promoter of BIRC5,and the dual-luciferase reporter gene assay was applied to detect the transcriptional activity of AP-4 on promoter of BIRC5.Results The number of apoptosis and the apoptosis rate in HEC-1-A and RL95-2 cells of the si#1 group and si#2 group was significantly increased,while the survivin mRNA expression was decreased as compared with that of the control group (all P <0.05).A high-confidence AP-4 binding site existed in the BIRC5 promoter region,and si-lencing AP-4 suppressed the binding of AP-4 on the promoter region of BIRC5,and suppressed the transcriptional activity of AP-4 on promoter of BIRC5.Conclusions Silencing AP-4 could induce the apoptosis of endometrial cancer cells through re-ducing the AP-4 on survivin transcription activation,and thereby inducing apoptosis of tumor cells.