丙泊酚影响视网膜母细胞瘤细胞生长和干样特性的机制研究

Effect of Propofol on Growth and Stem Cell-like Features of Retinoblastoma Cells and the Mechanisms

目的探究丙泊酚影响视网膜母细胞瘤(retinoblastoma,RB)细胞生长和干样特性的调控机制。方法选取RB细胞系Y79为研究对象,利用CCK-8法进行浓度梯度实验确定后续实验浓度,利用克隆形成实验和流式细胞术分析丙泊酚处理对Y79细胞的生长率及凋亡率的影响,利用干细胞成球实验分析丙泊酚处理对Y79细胞干样特性的影响;Western印迹检测分析丙泊酚处理后干细胞特性蛋白S0X2、OCT4、CD44以及AMPKα1、p-Nrf2、Nrf2、HO-1蛋白的表达水平。结果CCK-8梯度浓度试验发现丙泊酚浓度从5μg/ml开始对Y79细胞活性产生了明敁抑制作用(P<0.05),后续以2.5、5、10μg/ml为试验浓度;与无药处理的对照组相比,5、10μg/ml丙泊酚干预后克隆形成率、干细胞成球率、成球直径、S0X2、0CT4、CD44、AMPKα1、p-Nrf2/Nrf2、HO-1表达水显著降低,凋亡率显著升高(P<0.05)。结论丙泊酚可以在体外抑制AMPK/Nrf2/HO-1信号通路激活,降低RB细胞抗氧化应激能力,抑制其生长及干细胞特性的维持而发挥抗癌作用.

Objective To explore the mechanism by which propofol influences the growth and stem cell-like features of retinoblastoma(RB)cells.Methods RB cell line Y79 cells were selected as tlie lesearch objects.Concentration gradient experiment was conducted with CCK-8 method to determine the concentrations of subsequent experiments.The effects of propofol treatment on growth and apoptosis rates of Y79 cells were analyzed by clone formation assay and flow cytometry.The effects of propofol treatment on Y79 stem cell-like features were analyzed by stem cells pelletization experiment.The expression levels of stem cells features(S0X2,OCT4,CD44),AMPKα1,p-Nrf2,Nrf2 and HO-1 proteins after propofol treatment were detected and analyzed by Western blotting.Results It was found by concentration gradient experiment that thatpropofol(not lower than 5μg/mL)exerted significant inhibitory effects on the activity of Y79 cells(P<0.05),and the concentration values(2.5,5,10μg/mL)were selected for subsequent experiment.Compared with control group without drug treatment,colony formal ion rate,pelletization rate of stem cells,pelletization size,expression levels of S0X2,0CT4,CD44,AM PKα1,p-Nif2/Nrf2 and HO-1 were significantly decreased,while apoptosis rate was significantly increased after 5 and 10μg/mL propofol intervention(P<0.05).Conclusion Propofol can inhibit the activation of AMPK/Nrf2/HO-1 signaling pathwayin vitro,reduce anti-oxidative stress ability of RB cells,inhibit their growth and maintenance of slem cell-like features,and thus playing anti-cancer roles.