亚硒酸钠通过增加ROS抑制PI3K/AKT通路改善肺癌PC-9/GR细胞对吉非替尼的耐药性  

Sodium selenite reverses the resistance of lung cancer PC-9/GR cells to gefitinib by increasing ROS and inhibiting PI3K/AKT pathway

作　　者：李艳梅 马琳 刘单 邓述恺 LI Yanmei;MA Lin;LIU Dan;DENG Shukai(Department of Respiratory and Critical Care Medicine,Affiliated Hospital of Southwest Medical University,Sichuan Luzhou 646000,China)

机构地区：[1]西南医科大学附属医院呼吸与危重症医学科,四川泸州646000

出　　处：《现代肿瘤医学》2024年第4期636-641,共6页Journal of Modern Oncology

摘　　要：目的:探讨亚硒酸钠对肺癌耐药细胞PC-9/GR增殖、凋亡的影响,研究其是否能改善吉非替尼耐药及可能机制。方法:不同浓度(0、5、10、20、40μmol/L)亚硒酸钠处理细胞24、36、48 h后,CCK-8法检测亚硒酸钠对细胞增殖的影响并选择合适浓度用于后续实验。不同浓度(0、2.5、5、10、20、40μmol/L)吉非替尼单药及联合亚硒酸钠(6μmol/L)处理细胞48 h,分别计算半数抑制浓度(IC50),得出亚硒酸钠对PC-9/GR逆转倍数;流式细胞术检测各组细胞凋亡;DCFH-DA法检测细胞内ROS水平;Western blot实验检测各组细胞中p-PI3K、p-AKT、Bax、Bcl-2表达水平。结果:随亚硒酸钠浓度升高及作用时间延长,细胞增殖抑制率逐渐升高(P<0.05);单药吉非替尼组的IC50值为16.051μmol/L,联合亚硒酸钠后IC50值为5.406μmol/L,表明亚硒酸钠能够逆转吉非替尼的耐药,其逆转耐药倍数为2.969倍。与吉非替尼和亚硒酸钠单药组相比,联合治疗组的细胞凋亡率显著升高,ROS水平及Bax蛋白表达上调,p-PI3K、p-AKT、Bcl-2表达下调,经N-乙酰半胱氨酸(NAC)消除ROS后抵消了亚硒酸钠对PI3K/AKT通路的抑制作用(P<0.05)。结论:亚硒酸钠联合吉非替尼能提高肺腺癌耐药细胞的敏感性,抑制细胞的增殖,同时诱导细胞凋亡,这一过程可能通过ROS依赖性下调PI3K/AKT通路的表达实现。Objective:To investigate the effects of sodium selenite on the proliferation and apoptosis of lung cancer resistant cells PC-9/GR,and to study whether sodium selenite can improve the resistance to gefitinib and its potential mechanism.Methods:PC-9/GR cells were treated with different concentrations of sodium selenite(0,5,10,20,40μmol/L)for 24,36,48 hours.The proliferation of PC-9/GR cells was detected by CCK-8 assay,and the appropriate concentration of sodium selenite was selected for subsequent experiments.Cells were treated with different concentrations(0,2.5,5,10,20,40μmol/L)of gefitinib alone or in combination with sodium selenite(6μmol/L)for 48 hours.The median inhibitory concentration(IC 50)of each time group and the reversal ratio of sodium selenite to PC-9/GR were calculated.Apoptosis was detected by flow cytometry.ROS level was detected by DCFH-DA.The expression levels of p-PI3K,p-AKT,Bax and Bcl-2 were detected by Western blot.Results:With the increase of sodium selenite concentration and the longer treatment time,the cell proliferation inhibition rate increased(P<0.05).The IC 50 of gefitinib alone was 16.051μmol/L,while in combination with sodium selenite was 5.406μmol/L.Indicating that sodium selenite could reverse the drug resistance of gefitinib by 2.969 times.Compared with the monotherapy groups,the apoptosis rate of combination group significantly increased.The levels of ROS and the expression of Bax protein were upregulated,while the expression of p-PI3K,p-AKT,and Bcl-2 were downregulated.After eliminating ROS with N-acetylcysteine(NAC),the inhibitory effect of sodium selenite on the PI3K/AKT pathway was counteracted(P<0.05).Conclusion:Sodium selenite combined with gefitinib can enhance the sensitivity of gefitinib-resistant lung adenocarcinoma cells,inhibit cell proliferation,and induce cell apoptosis.This process may be achieved by downregulating the expression of the PI3K/AKT pathway through ROS-dependent manner.

关 键 词：肺癌 亚硒酸钠 活性氧(ROS) EGFR-TKIs耐药 PI3K/AKT通路 

分 类 号：R734.2[医药卫生—肿瘤]