中国肺癌杂志
中國肺癌雜誌
중국폐암잡지
Chinese Journal of Lung Cancer
2015年
10期
599-609
,共11页
张鑫宇%刘皈阳%刘浩%马涛
張鑫宇%劉皈暘%劉浩%馬濤
장흠우%류귀양%류호%마도
T790M%H1975%等效线图法%雷公藤甲素%吉非替尼%凋亡
T790M%H1975%等效線圖法%雷公籐甲素%吉非替尼%凋亡
T790M%H1975%등효선도법%뢰공등갑소%길비체니%조망
T790M%H1975%Isobolograms%Triptolide%Getfinib%Apoptosis
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。
揹景與目的錶皮生長因子受體（epidermal growth factor receptor, EGFR）酪氨痠激酶抑製劑（tyrosine kinase inhibitors, TKIs）被用于治療進展性晚期非小細胞肺癌（non-small cell lung cancer, NSCLC），然而最初接受TKIs治療有反應的患者，大都會在10箇月左右齣現穫得性耐藥。報告稱EGFR基因T790M的突變是產生穫得性耐藥的主要原因，比例約佔50%。本研究旨在探索雷公籐甲素（triptolide, TP）和吉非替尼序貫應用對肺腺癌細胞H1975細胞增殖和凋亡通路的作用。方法 MTT法檢測細胞的增殖。等效線圖法和聯閤指數（combination index, CI）法評估雷公籐甲素和吉非替尼序貫作用的效價。流式細胞術檢測細胞凋亡和週期分佈，Hoechest 33258染色法檢測凋亡形態。化學比色髮光法檢測Caspases的活性。結果等效線圖法和聯閤指數法均顯示雷公籐甲素序貫吉非替尼組較其他序貫作用組明顯抑製瞭細胞增殖，增加瞭細胞的凋亡。細胞週期分佈實驗結果顯示與吉非替尼序貫雷公籐甲素組主要把細胞抑製在G0/G1期相比較，雷公籐甲素序貫吉非替尼組主要把細胞抑製在G2/M期。在肺腺癌H1975中，所有序貫模型組都主要通過活化Caspase-9/Caspase-3來誘導激活細胞凋亡通路。結論先用雷公籐甲素再用吉非替尼治療模式可能是剋服T790M突變耐藥的一箇新選擇。
배경여목적표피생장인자수체（epidermal growth factor receptor, EGFR）락안산격매억제제（tyrosine kinase inhibitors, TKIs）피용우치료진전성만기비소세포폐암（non-small cell lung cancer, NSCLC），연이최초접수TKIs치료유반응적환자，대도회재10개월좌우출현획득성내약。보고칭EGFR기인T790M적돌변시산생획득성내약적주요원인，비례약점50%。본연구지재탐색뢰공등갑소（triptolide, TP）화길비체니서관응용대폐선암세포H1975세포증식화조망통로적작용。방법 MTT법검측세포적증식。등효선도법화연합지수（combination index, CI）법평고뢰공등갑소화길비체니서관작용적효개。류식세포술검측세포조망화주기분포，Hoechest 33258염색법검측조망형태。화학비색발광법검측Caspases적활성。결과등효선도법화연합지수법균현시뢰공등갑소서관길비체니조교기타서관작용조명현억제료세포증식，증가료세포적조망。세포주기분포실험결과현시여길비체니서관뢰공등갑소조주요파세포억제재G0/G1기상비교，뢰공등갑소서관길비체니조주요파세포억제재G2/M기。재폐선암H1975중，소유서관모형조도주요통과활화Caspase-9/Caspase-3래유도격활세포조망통로。결론선용뢰공등갑소재용길비체니치료모식가능시극복T790M돌변내약적일개신선택。
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 atfer a progression-free period of about 10 months. hTe percentage of T790M in TKI acquired-resistant patients in most studies is around 50%. hTe aim of this study is to assess the effects of the sequential administration of triptolide and getfinib on cell proliferation and apoptosis of lung adenocarcinoma cell H1975. Methods A MTT assay was used to measure cell proliferation. hTe potency of the sequential ad-ministration of triptolide and getfinib were determined by isobolograms and combination index (CI). Cell apoptosis and cycle distribution were detected by lfow cytometry. hTe Hoechst 33258 method was used to observe the apoptotic morphology. Chemical colorimetric luminescence was used to measure the caspase activity. Results hTe results of isobolograms and CI showed that the sequential administration of triptolide following getfinib remarkably inhibited cell proliferation and cell apop-tosis compared with other sequential administration models. hTe cycle distribution results indicated that sequential triptolide administration following getfinib blocked the cells in the G2/M phase but not in the G0/G1 phase. hTe 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 hTe triptolide administration following getfinib might be a new therapeutic strategy for lung cancer with T790M mutation atfer having EGFR-TKIs resistance.
