Objective:The aim of the study was to explore the effect of STAT5 silenced by siRNA on proliferation,apoptosis and invasion of esophageal carcinoma cell line EC9706.Methods:The siRNA vectors aiming to STAT5 gene were ...Objective:The aim of the study was to explore the effect of STAT5 silenced by siRNA on proliferation,apoptosis and invasion of esophageal carcinoma cell line EC9706.Methods:The siRNA vectors aiming to STAT5 gene were constructed.STAT5 siRNA was transfected into EC9706 cells by Lipofectamine TM 2000.Changes of STAT5,Bcl-2 and Cyclin D1 were analyzed by Western blot and RT-PCR.Effect of STAT5 siRNA on EC9706 cells proliferation was determined by MTT.Effects of STAT5 siRNA on EC9706 cells cycle and apoptosis were detected by the flow cytometry.Boyden chamber was used to evaluate the invasion and metastasis capabilities of EC9706 cells.Results:The double strands oligonucleotide of siRNA aiming to STAT5 was successfully cloned into the pRNAT-U6.1 vector,and the target sequence was coincide with the design.RT-PCR and Western blotting detection demonstrated that the expression levels of STAT5,Bcl-2 and Cyclin D1 genes were obviously decreased in EC9706 cells transfected with STAT5 targeting siRNA expression vectors.STAT5 siRNA could suppress the proliferation of EC9706 cells.The proportions of S and G2/M periods frequency were significantly decreased (P < 0.05),and the proportion of G0/G1 period frequency was significantly increased (P < 0.05).The average amount of cells penetrating Matrigel was significantly decreased (P < 0.05).Silencing the STAT5 induced the apoptosis of esophageal carcinoma cell line EC9706 (P < 0.01).Conclusion:STAT5 silenced by siRNA of esophageal carcinoma cell line EC9706 could induce the apoptosis.And it could suppress the proliferation,invasion and metastasis of tumor cells.展开更多
文摘Objective:The aim of the study was to explore the effect of STAT5 silenced by siRNA on proliferation,apoptosis and invasion of esophageal carcinoma cell line EC9706.Methods:The siRNA vectors aiming to STAT5 gene were constructed.STAT5 siRNA was transfected into EC9706 cells by Lipofectamine TM 2000.Changes of STAT5,Bcl-2 and Cyclin D1 were analyzed by Western blot and RT-PCR.Effect of STAT5 siRNA on EC9706 cells proliferation was determined by MTT.Effects of STAT5 siRNA on EC9706 cells cycle and apoptosis were detected by the flow cytometry.Boyden chamber was used to evaluate the invasion and metastasis capabilities of EC9706 cells.Results:The double strands oligonucleotide of siRNA aiming to STAT5 was successfully cloned into the pRNAT-U6.1 vector,and the target sequence was coincide with the design.RT-PCR and Western blotting detection demonstrated that the expression levels of STAT5,Bcl-2 and Cyclin D1 genes were obviously decreased in EC9706 cells transfected with STAT5 targeting siRNA expression vectors.STAT5 siRNA could suppress the proliferation of EC9706 cells.The proportions of S and G2/M periods frequency were significantly decreased (P < 0.05),and the proportion of G0/G1 period frequency was significantly increased (P < 0.05).The average amount of cells penetrating Matrigel was significantly decreased (P < 0.05).Silencing the STAT5 induced the apoptosis of esophageal carcinoma cell line EC9706 (P < 0.01).Conclusion:STAT5 silenced by siRNA of esophageal carcinoma cell line EC9706 could induce the apoptosis.And it could suppress the proliferation,invasion and metastasis of tumor cells.
