摘要
目的探讨吉非替尼和依维莫司对吉非替尼耐药鼻咽癌细胞的作用效果及分子作用机制。方法吉非替尼、依维莫司单独或联合作用人鼻咽癌细胞株HONE1后,采用噻唑蓝(MTT)法检测细胞的增殖抑制,流式细胞术检测细胞凋亡率及周期分布,Western blot检测细胞磷酸化丝氨酸苏氨酸蛋白激酶(p-AKT)、磷酸化核糖体S6蛋白激酶(pS6K)的表达水平。结果吉非替尼和依维莫司均呈浓度依赖性抑制HONE1细胞增殖,吉非替尼的半数抑制浓度(IC50)为17.92μmol·L-1,依维莫司的IC50为2.46 nmol·L-1,但二者联合作用效果与单药相比并未表现出明显的优势(P>0.05);吉非替尼对p-AKT抑制作用不明显,依维莫司可下调p-S6K的表达,同时上调了p-AKT的表达,联合用药对pS6K和p-AKT表达的抑制作用并不明显强于单药。结论吉非替尼和依维莫司联合作用于吉非替尼耐药鼻咽癌细胞株HONE1没有表现出协同效应,表皮生长因子受体/丝氨酸苏氨酸蛋白激酶(EGFR/AKT)信号通路与雷帕霉素蛋白(mTOR)信号通路在鼻咽癌细胞中的关系有待进一步研究。
Objective To investigate the synergistic effect and molecular mechanism of the epidermal growth factor receptor （EGFR） inhibitor gefitinib and the mammalian target of rapamycin （roTOR） inhibitor everolimus on nasopharyngeal carcinoma resistant to gefitinib in vitro. Methods Human nasopharyngeal carcinoma cell line HONE1 were incubated with gefitinib and everolimus alone and in combination, the effect of cell growth inhibition was measured by MT＇F assay. Flow cytometry was applied to assess the cell cycle and apoptosis. Western blot was used to detect the expression of phosphorylated levels of protein serine threonine kinases （p-AKT） and ribosomal $6 kinases （p-S6K） in cells. Results The effects of cell growth inhibition by gefitinib or everolimus separately were dose-dependent in the HONEI cell line. The ICso value for gefitinib was 17.92 i^mol ~ L-1 , and that for everolimus was 2.46 nmol ~ L-I. However, the combination of two agents did not show a obvious advantage over the drug used alone（ P〉0.05 ）. The inhibition of p-AKT expression was not remarkable in the HONE1 cell line after gefitinib treatment. Everolimus was revealed to down-regulate the expression of po-S6K and up-regulate the level of p-AKT in the HONE1 cell line. Moreover, no stronger inhibitory effect on p-S6K and p-AKT expression by combined agents was found. Conclusion Combined treatment of gefitinib and everolimus doesn＇ t show a synergistic effect against nasopharyngeal carcinoma cell line HONE1 resistant to gefitinib. Further research is in need to investigate the relationship between the EGFR/ AKT signaling pathway and the roTOR signaling pathway in nasopharyngeal carcinoma cells.
出处
《医药导报》
CAS
北大核心
2014年第1期43-48,共6页
Herald of Medicine
基金
吴阶平医学基金资助项目(320.6700.09065)
国家自然科学基金资助项目(81272491)
关键词
吉非替尼
依维莫司
癌
鼻咽
联合用药
Gefitinib
Everolimus
Carcinoma, nasopharyngeal
Combination therapy
