Rab11通过NF-κB信号通路调控膀胱癌细胞的增殖和侵袭

摘要

Objective:To investigate the molecular mechanism of Rab11 regulation on proliferation and invasion of bladder cancer cell.Methods:The endogenous expression of Rab11 in bladder cancer BIU-87,T24,RT4 and 5637 cell lines was detected by Western blot.In BIU-87(Rab11 low expression)and T24(Rab11 overexpression)cell lines,Rab11 overexpressing plasmids and Rab11 siRNA were transfected.Luciferase assay was used to detect the ac-tivity of NF-κB signaling pathway in transfected cells.The expression of NF-κB signaling related protein p-IκB was analyzed by Western blot technique. BIU -87 cell lines transfected with Rab11 overexpressing plasmids were treated with NF-κB inhibitor Bay 11-7082. The protein expression of cell cycle related factors cyclin D1 and inva-sion related factor MMP9 were detected by Western blot.Results:In the BIU-87 cell lines transfected with Rab11 overexpression plasmid,the activity of NF-κB increased,and the expression of p-IκB increased.After knockdown of Rab11,the activity of NF-κB was inhibited,and the expression of p-IκB was inhibited.The activity of p-IκB was positively correlated with the activity of NF-κB signaling pathway.Inhibitor Bay 11-7082 of NF-κB blocked the process of Rab11 induced upregulation of cyclin D1 and MMP9.Conclusion:Rab11 regulates the proliferation and in-vasion of bladder cancer cells through the NF-κB signaling pathway.%目的:探讨Rab11调控膀胱癌细胞增殖和侵袭的分子机制.方法:用Western blot方法检测膀胱癌BIU-87、T24、RT4、5637细胞系中Rab11的内源表达量.在BIU-87(Rab11低表达)和T24(Rab11高表达)细胞系中分别转染Rab11过表达质粒和Rab11 siRNA.用荧光素酶报告实验检测转染后细胞内的NF-κB信号通路的活性,通过Western blot技术分析与NF-κB信号通路相关蛋白p-IκB的变化情况.用NF-κB抑制剂Bay 11-7082处理转染Rab11过表达质粒的BIU-87细胞系,通过Western blot检测细胞系中细胞周期相关因子cyclin D1和侵袭相关因子MMP9蛋白的变化.结果:转染Rab11过表达质粒的BIU-87细胞系中NF-κB活性水平提高,p-IκB表达量显著提高;而敲除Rab11后NF-κB活性受到抑制,p-IκB的表达受到抑制,而p-IκB与NF-κB信号通路的活性呈正相关.NF-κB抑制剂Bay 11-7082阻遏了Rab11诱导cyc-lin D1和MMP9表达上调的过程.结论:Rab11通过NF-κB信号通路调控膀胱癌细胞的增殖和侵袭.