中国肺癌杂志
中國肺癌雜誌
중국폐암잡지
CHINESE JOURNAL OF LUNG CANCER
2014年
11期
778-782
,共5页
肺肿瘤%热休克蛋白90抑制剂%表皮生长因子受体酪氨酸激酶抑制剂%增殖%凋亡
肺腫瘤%熱休剋蛋白90抑製劑%錶皮生長因子受體酪氨痠激酶抑製劑%增殖%凋亡
폐종류%열휴극단백90억제제%표피생장인자수체락안산격매억제제%증식%조망
Lung neoplasms%HSP90 inhibitors%EGFR-TKI%Proliferation%Apoptosis
背景与目的表皮生长因子受体酪氨酸激酶抑制剂（epidermal growth factor receptor-tyrosine kinase inhibi-tor, EGFR-TKI）在非小细胞肺癌（non-small cell lung cancer, NSCLC）患者的临床治疗中产生的原发性及获得性耐药限制了其临床应用，需要探索新的策略或方法来克服这个问题。最近有文献报道认为热休克蛋白90（heat shock protein 90, HSP90）抑制剂能从多种途径和环节发挥抗肿瘤作用，这为解决NSCLC对EGFR-TKI的耐药提供了新的思路。本研究通过观察HSP90抑制剂17-DMAG对EGFR-TKI分别原发性及获得性耐药的NSCLC细胞株A549和H1975的作用，旨在探讨它对细胞增殖、凋亡与EGFR蛋白表达的影响及其可能的机制。方法以不同浓度的17-DMAG分别作用于A549和H1975细胞株24 h、48 h、72 h，应用四甲基偶氮唑蓝（MTT）比色法检测细胞增殖；作用48 h后，应用流式细胞术PI单染法检测细胞凋亡，并应用Western blot检测细胞HSP90及EGFR蛋白表达水平。结果17-DMAG在不同药物浓度和作用时间对A549和H1975细胞的增殖抑制率差异均有统计学意义（P<0.01），且呈时间和剂量依赖性；两种细胞不同药物浓度组和空白对照组之间的凋亡率差异均有统计学意义（P<0.01），且呈剂量依赖性；17-DMAG作用48 h后，A549细胞的EGFR/GADPH和HSP90/GADPH及H1975细胞的HSP90/GADPH在不同药物浓度组和空白对照组之间的灰度比值差异均无统计学意义（P>0.05），而H1975细胞的EGFR/GADPH在不同药物浓度组和空白对照组之间的灰度比值差异均有统计学意义（P<0.01）。结论17-DMAG对NSCLC细胞株A549和H1975均具有抑制增殖及促进凋亡作用，且它能降低突变型EGFR的蛋白表达水平，而对野生型EGFR的蛋白表达无明显影响。本研究为EGFR-TKI耐药的非小细胞肺癌提供了新的治疗策略。
揹景與目的錶皮生長因子受體酪氨痠激酶抑製劑（epidermal growth factor receptor-tyrosine kinase inhibi-tor, EGFR-TKI）在非小細胞肺癌（non-small cell lung cancer, NSCLC）患者的臨床治療中產生的原髮性及穫得性耐藥限製瞭其臨床應用，需要探索新的策略或方法來剋服這箇問題。最近有文獻報道認為熱休剋蛋白90（heat shock protein 90, HSP90）抑製劑能從多種途徑和環節髮揮抗腫瘤作用，這為解決NSCLC對EGFR-TKI的耐藥提供瞭新的思路。本研究通過觀察HSP90抑製劑17-DMAG對EGFR-TKI分彆原髮性及穫得性耐藥的NSCLC細胞株A549和H1975的作用，旨在探討它對細胞增殖、凋亡與EGFR蛋白錶達的影響及其可能的機製。方法以不同濃度的17-DMAG分彆作用于A549和H1975細胞株24 h、48 h、72 h，應用四甲基偶氮唑藍（MTT）比色法檢測細胞增殖；作用48 h後，應用流式細胞術PI單染法檢測細胞凋亡，併應用Western blot檢測細胞HSP90及EGFR蛋白錶達水平。結果17-DMAG在不同藥物濃度和作用時間對A549和H1975細胞的增殖抑製率差異均有統計學意義（P<0.01），且呈時間和劑量依賴性；兩種細胞不同藥物濃度組和空白對照組之間的凋亡率差異均有統計學意義（P<0.01），且呈劑量依賴性；17-DMAG作用48 h後，A549細胞的EGFR/GADPH和HSP90/GADPH及H1975細胞的HSP90/GADPH在不同藥物濃度組和空白對照組之間的灰度比值差異均無統計學意義（P>0.05），而H1975細胞的EGFR/GADPH在不同藥物濃度組和空白對照組之間的灰度比值差異均有統計學意義（P<0.01）。結論17-DMAG對NSCLC細胞株A549和H1975均具有抑製增殖及促進凋亡作用，且它能降低突變型EGFR的蛋白錶達水平，而對野生型EGFR的蛋白錶達無明顯影響。本研究為EGFR-TKI耐藥的非小細胞肺癌提供瞭新的治療策略。
배경여목적표피생장인자수체락안산격매억제제（epidermal growth factor receptor-tyrosine kinase inhibi-tor, EGFR-TKI）재비소세포폐암（non-small cell lung cancer, NSCLC）환자적림상치료중산생적원발성급획득성내약한제료기림상응용，수요탐색신적책략혹방법래극복저개문제。최근유문헌보도인위열휴극단백90（heat shock protein 90, HSP90）억제제능종다충도경화배절발휘항종류작용，저위해결NSCLC대EGFR-TKI적내약제공료신적사로。본연구통과관찰HSP90억제제17-DMAG대EGFR-TKI분별원발성급획득성내약적NSCLC세포주A549화H1975적작용，지재탐토타대세포증식、조망여EGFR단백표체적영향급기가능적궤제。방법이불동농도적17-DMAG분별작용우A549화H1975세포주24 h、48 h、72 h，응용사갑기우담서람（MTT）비색법검측세포증식；작용48 h후，응용류식세포술PI단염법검측세포조망，병응용Western blot검측세포HSP90급EGFR단백표체수평。결과17-DMAG재불동약물농도화작용시간대A549화H1975세포적증식억제솔차이균유통계학의의（P<0.01），차정시간화제량의뢰성；량충세포불동약물농도조화공백대조조지간적조망솔차이균유통계학의의（P<0.01），차정제량의뢰성；17-DMAG작용48 h후，A549세포적EGFR/GADPH화HSP90/GADPH급H1975세포적HSP90/GADPH재불동약물농도조화공백대조조지간적회도비치차이균무통계학의의（P>0.05），이H1975세포적EGFR/GADPH재불동약물농도조화공백대조조지간적회도비치차이균유통계학의의（P<0.01）。결론17-DMAG대NSCLC세포주A549화H1975균구유억제증식급촉진조망작용，차타능강저돌변형EGFR적단백표체수평，이대야생형EGFR적단백표체무명현영향。본연구위EGFR-TKI내약적비소세포폐암제공료신적치료책략。
Background and objective In the clinical treatment of patients with non-small cell lung cancer (NSCLC), the primary and acquired resistance of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) limits its clinical application, this need to explore new strategy or method to overcome this problem. Recently, some literatures have indicated that the antitumor role of heat shock protein 90 (HSP90) inhibitors by a variety of pathways may provide new strategy for resolving this problem. In this study, we examined the effect of 17-DMAG on NSCLC cell lines A549 and H1975 which were primary and acquired resistant to EGFR-TKI respectively, the purpose was to explore its inlfuence on cell prolifera-tion, apoptosis and the expression of EGFR in vitro as well as possible mechanism. Methods Atfer A549 and H1975 cell lines were treated with different concentrations of 17-DMAG respectively, the inhibitory rate of cell proliferation was measured by MTT assay in 24 h, 48 h and 72 h. We investigated the effect of 17-DMAG on the cell apoptosis with lfow cytometry and the ex-pression of HSP90 and EGFR with Western blot atfer treated with 17-DMAG for 48 h. Results Atfer treated with 17-DMAG, the inhibitory rate of different concentrations and time groups was signiifcant (P<0.01), and the effect was in time-and dose-dependent manner;the apoptosis rate of both two cell lines in all treated groups were signiifcantly higher than control group (P<0.01), and the effect was in dose-dependent manner. By Western blot analysis, there was no signiifcant difference between all treated groups and control group for the expression of both HSP90 and EGFR protein in A549 cell line and HSP90 protein in H1975 cell line atfer exposed to 17-DMAG for 48 h (P>0.05), while the difference was signiifcant for the expression of EGFR protein in H1975 cell line (P<0.01). Conclusion 17-DMAG inhibited the proliferation of NSCLC cell lines A549 and H1975 and also induced apoptosis of both cell lines. It down-regulated the expression of mutant EGFR protein while this phe-nomenon was not observed in EGFR-wild type cell line. hTis suggested that the mechanism maybe different between A549 and H1975 cell lines with different genetic backgroud. Our study provided new strategy for treatment with NSCLC being resistant to EGFR-TKI.
