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Expression and regulaition of survivin shRNA interference on nuclear factorκB in esophageal ECA109 cells
LI Xiu-mei CHEN Yan JIANG Xiao-fang LI HuiGUO Qiong LI Hui-wu SHEN Hong-ming*
Acta Anatomica Sinica ›› 2015, Vol. 46 ›› Issue (4) : 514-520.
Expression and regulaition of survivin shRNA interference on nuclear factorκB in esophageal ECA109 cells
Objective As the effect of Survivin, a gene expression regulatory factor, on the nuclear factor (NF)-kappa B gene transcription and protein activity remains unknown, this subject aims to explore the possible regulation mechanism of Survivin and NF-kappa B. Methods Survivin shRNA interference technology was applied to liposome transfect esophageal ECA109 cells, detect Survivin silence result, choose the best transfection concentration; Half quantitative RT-PCR was applied to detect genes expression of NF-kappa B and upstream regulatory factors inhibitor of NF-kappa B kinase alpha(IKK alpha), IKK beta in the same pathway; Western blotting was used to detect protein expression of the NF-kappa B. Flow cytometry was used to detect the proliferation and apoptosis of cell lines index changes after ECA109 esophageal cells transfection. Results After survivin shRNA transfection in esophageal ECA109 cells, survivin mRNA and protein expression were obviously down-regulated. NF-kappa B mRNA expression had no obvious difference, however the phosphorylated proteins level of NF-kappaB reduced and its upstream factor IKK alpha, IKK beta mRNA express reduced as well. Esophageal cell ECA109 apoptosis increased significantly, the cells ratio increased in G2 phase but decreased in S phase, and the cell cycle was blocked. Conclusion Survivin regulates NF-kappa B activation by down-regulation of IKK alpha and IKK beta mRNA expression, suggesting that survivin may be a new gene regulatory factor in the tumor signal control network.
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