摘要
背景与目的表皮生长因子受体(epidermal growth factor receptor,EGFR)酪氨酸激酶抑制剂(tyrosine kinase inhibitors,TKIs)被用于治疗进展性晚期非小细胞肺癌(non-small cell lung cancer,NSCLC),然而最初接受TKIs治疗有反应的患者,大都会在10个月左右出现获得性耐药。报告称EGFR基因T790M的突变是产生获得性耐药的主要原因,比例约占50%。本研究旨在探索雷公藤甲素(triptolide,TP)和吉非替尼序贯应用对肺腺癌细胞H1975细胞增殖和凋亡通路的作用。方法 MTT法检测细胞的增殖。等效线图法和联合指数(combination index,CI)法评估雷公藤甲素和吉非替尼序贯作用的效价。流式细胞术检测细胞凋亡和周期分布,Hoechest 33258染色法检测凋亡形态。化学比色发光法检测Caspases的活性。结果等效线图法和联合指数法均显示雷公藤甲素序贯吉非替尼组较其他序贯作用组明显抑制了细胞增殖,增加了细胞的凋亡。细胞周期分布实验结果显示与吉非替尼序贯雷公藤甲素组主要把细胞抑制在G0/G1期相比较,雷公藤甲素序贯吉非替尼组主要把细胞抑制在G2/M期。在肺腺癌H1975中,所有序贯模型组都主要通过活化Caspase-9/Caspase-3来诱导激活细胞凋亡通路。结论先用雷公藤甲素再用吉非替尼治疗模式可能是克服T790M突变耐药的一个新选择。
Background and objective Epidermal growth factor receptor （EGFR） tyrosine kinase inhibitors （TKIs） show promising therapeutic effects in patients with advanced non-small cell lung cancer （NSCLC）. However, despite an initial response to TKIs treatment among responsive patients, most inevitably acquire resistance after a progression-free period of about 10 months, q-he percentage of T790M in TKI acquired-resistant patients in most studies is around 50%. The aim of this study is to assess the effects of the sequential administration of triptolide and geftinib on cell proliferation and apoptosis of lung adenocarcinoma cell H1975. Methods A M＇IT assay was used to measure cell proliferation. The potency of the sequential ad- ministration of triptohde and geftinib were determined by isobolograms and combination index （CI）. Ceil apoptosis and cycle distribution were detected by flow cytometry. The Hoechst 33258 method was used to observe the apoptotic morphology. Chemical colorimetric luminescence was used to measure the caspase activity. Results The results ofisobolograms and CI showed that the sequential administration of triptolide following geftinib remarkably inhibited cell proliferation and cell apop- tosis compared with other sequential administration models. The cycle distribution results indicated that sequential triptolide administration following geftinib blocked the cells in the G2/M phase but not in the G0/G1 phase. The activation of the Cas- pase-9/Caspase-3 cascade was mainly involved in the apoptotic pathway of lung adenocarcinoma cell H1975 in all sequential administration models. Conclusion The triptolide administration following geftinib might be a new therapeutic strategy for lung cancer with T790M mutation after having EGFR-TKIs resistance.
出处
《中国肺癌杂志》
CAS
CSCD
北大核心
2015年第10期599-609,共11页
Chinese Journal of Lung Cancer
基金
蚌埠医学院科研课题项目(No.Byky1324)资助~~
