【摘要】 目的:探讨在鼻咽癌细胞中顺铂(DDP)是否能通过改变表皮生长因子受体(EGFR)和蛋白激酶B(AKT)的磷酸化蛋白水平以及对吉非替尼疗效的影响,寻找能够预测两者联合效应的分子指标。方法:MTT法分别测定CNE1、CNE2和SUNE1细胞中单药DDP和吉非替尼以及两药三种顺序联合的生长抑制率。蛋白质印迹法检测DDP、吉非替尼不同浓度对CNE1、CNE2细胞内EG-FR、p-EGFR、AKT以及p-AKT表达的影响。结果:DDP和吉非替尼单药在一定作用浓度内对CNE1、CNE2和SUNE1细胞生长抑制作用呈时间和剂量依赖性。随吉非替尼浓度增加,可以使CNE1、CNE2细胞中的p-EGFR、p-AKT表达下调。在CNE1细胞中,DDP与吉非替尼相互拮抗,且先用DDP后用吉非替尼较先用吉非替尼后用DDP拮抗效应更明显,P=0.039;DDP可以诱导CNE1细胞的p-EGFR和p-AKT表达水平下调。而在CNE2细胞中,DDP与吉非替尼相互协同,DDP可以诱导p-EGFR和p-AKT表达水平上调。结论:DDP与吉非替尼联合在CNE1和CNE2细胞中分别显示拮抗和协同效应。DDP对吉非替尼效应的影响可能是通过改变p-EGFR和p-AKT蛋白水平来实现的。更多还原

【Abstract】 OBJECTIVE:To investigate that whether Cisplatin can affect the efficacy of Gefitinib through changes in protein expression levels of EGFR and AKT phosphorylation in nasopharyngeal carcinoma cells,and find the marker which can predict the effects of two-drug combination.METHODS:The cell proliferation inhibition rates of single agent of Cisplatin or Gefitinib and two drug combination in three soguehces were detected by MTT assay in CNE1,CNE2 and SUNE1 cell lines.The protein expression levels of EGFR,phospho-EGFR,AKT and phospho-AKT were detected by Western Blot analysis when CNE1 and CNE2 cell lines were exposed to Cisplatin or Gefitinib in different concentrations.RESULTS:In three cell lines,cell proliferation inhibition effect of Cisplatin or Gefitinib was time-and dose-dependent in a certain range of concentrations.With the increasing concentration of Gefitinib exposure,phospho-EGFR or phospho-AKT were down-regulated in CNE1 and CNE2 cell lines.With the increasing concentration of Cisplatin exposure,phospho-EGFR or phospho-AKT were down-regulated in CNE1cell in which antagonistic interaction between Cisplatin and Gefitinib were observed.In addition,the treatment of Cisplatin followed by Gefitinib showed greater antagonistic interaction than that of Gefitinib followed by Cisplatin in CNE1(P=0.039).With the increasing concentration of Cisplatin exposure,phospho-EGFR or phospho-AKT was up-regulated in CNE2 cell in which synergistic interaction between Cisplatin and Gefitinib were observed.CONCLUSIONS:Cisplatin combined with Gefitinib can result in antagonism and synergism in CNE1 and CNE2 cell line respectively.It may be through changes in protein expression levels of EGFR and AKT phosphorylation that Cisplatin can affect the efficacy of Gefitinib.更多还原