过表达NFKBIZ对膀胱癌细胞周期及凋亡的影响

Effect of NFKBIZ gene on cell cycle and apoptosis in bladder cancer cells

目的探究NFKBIZ基因对人膀胱癌细胞株T24及5637细胞周期及凋亡的影响,为膀胱肿瘤的发生、发展机制及其诊治提供理论依据。方法体外培养膀胱癌细胞T24及5637,利用慢病毒构建NFKBIZ过表达的稳定转染细胞株T24和5637,使用CCK-8实验和平板克隆实验探究过表达NFKBIZ对细胞增殖能力的影响,用Western blot检测过表达NFKBIZ后的凋亡相关蛋白及细胞周期蛋白的表达水平,用流式细胞仪检测过表达NFKBIZ后的细胞凋亡率和细胞周期的变化。结果过表达NFKBIZ后,细胞增殖能力降低,凋亡蛋白Bax和cleaved-caspase3表达水平升高,而抗凋亡蛋白Bcl-2表达水平显著下降,同时,细胞周期蛋白依赖性激酶2及细胞周期蛋白E1表达显著降低,而细胞周期负调节蛋白p27表达升高,且细胞凋亡率升高,细胞周期受到阻滞。结论过表达NFKBIZ后,膀胱癌细胞周期受到阻滞,并激活了细胞凋亡,这说明NFKBIZ可能在膀胱癌的发生、发展中发挥重要作用,为膀胱癌的分子靶向治疗提供了新的理论依据。

Objective To investigate the effect of NFKBIZ gene on the cycle and apoptosis of bladder cancer cells in T24 and 5637.Methods Bladder cancer T24 and 5637 cells were cultured in vitro.Lentivirus was used to up-regulate the expression of NFKBIZ in T24 and 5637 cells.CCK-8 experiment and plate cloning experiment were used to explore the effect of overexpression of NFKBIZ on cell proliferation.Western blot was used to detect the expression of apoptosis related proteins and cell cycle proteins after overexpression of NFKBIZ.Flow cytometry was used to detect the changes of apoptosis rate and cell cycle after overexpression of NFKBIZ.Results Overexpression of NFKBIZ reduced the ability of cell proliferation.Besides it increased the expression levels of apoptotic proteins Bax and cleaved-caspase 3,while the expression level of anti-apoptotic protein Bcl-2 decreased significantly.At the same time,the expression of cyclin CDK2 and cyclinE1 decreased significantly,while the expression of cell cycle negative regulator p27 increased.In addition,it also increased the rate of apoptosis and blocked the cell cycle.Conclusions Overexpression of NFKBIZ could block cell cycle and induce apoptosis.This indicates that NFKBIZ may play an important role in the occurrence,development,cell cycle and apoptosis of bladder cancer,and provides a new theoretical basis for molecular targeted therapy of bladder cancer.