17-丙烯胺基-17-去甲氧基格尔德霉素对人肺癌A549及H446细胞增殖和凋亡的影响
17-병희알기-17-거갑양기격이덕매소대인폐암A549급H446세포증식화조망적영향
Effects of 17-AAG on the proliferation and apoptosis of human lung cancer A549 and H446 cells
  • 万方数据
    中华结核和呼吸杂志 중화결핵화호흡잡지
    Chinese Journal of Tuberculosis and Respiratory Diseases
    2015年 4期 267-272 ,共6页

    牛奔%蔺景双%冯涛

    우분%린경쌍%풍도

    肺肿瘤%细胞系,肿瘤%细胞凋亡%细胞增殖 肺腫瘤%細胞繫,腫瘤%細胞凋亡%細胞增殖
    폐종류%세포계,종류%세포조망%세포증식
    Lung neoplasms%Cell line,tumor%Apoptosis%Cell proliferation
    目的 观察17-丙烯胺基-17-去甲氧基格尔德霉素(17-AAG)对人肺癌A549及H446细胞凋亡的影响,初步探讨其可能的作用机制.方法 实验组细胞分别使用浓度为50、100、200、400和500 nmol/L的17-AAG进行处理,以不经药物处理组为阴性对照,通过MTT法检测12、24、48及72 h4个时间段内17-AAG对A549及H446细胞增殖的影响.以不经药物处理组为阴性对照,通过流式细胞术检测浓度为100、200和400 nmol/L的17-AAG作用48 h后,对A549及H446细胞周期的影响及其促凋亡作用.以不经药物处理组为阴性对照,通过Western blot法检测浓度为100、200和400 nmol/L的17-AAG对A549及H446细胞内热休克蛋白(Hsp) 90相关蛋白(Hsp90、Hsp70、Akt和Her-2)和凋亡相关蛋白(Bcl-2和Bax)表达的影响.本研究中结果对比类计量资料以(-x)±s表示,采用Origin 8.0统计软件,多组间均数比较采用t检验,组间两两比较用单因素方差分析(One-way ANOVA).结果 17-AAG浓度为50～ 500 nmol/L时2个细胞株的增殖受抑制,呈显著的浓度依赖性.48 h时,17-AAG对A549及H446细胞的半抑制浓度(IC50)分别为(222±13)和(189 ±7) nmol/L;细胞周期实验结果发现,17-AAG可特异性地将A549及H446细胞株的细胞周期阻滞于G2/M期;凋亡实验结果显示,17-AAG可促进A549及H446细胞凋亡;17-AAG浓度为400 nmol/L处理48 h后,A549细胞株的细胞凋亡率为46.3％,H446细胞株的细胞凋亡率为56.9％,与未加药物处理的对照组相比(A549细胞株的细胞凋亡率为11.9％,H446细胞株的细胞凋亡率为6.9％)差异有统计学意义(P＜0.01);经17-AAG处理48 h后,2个细胞株中Hsp90信号通路的相关蛋白均发生了相同趋势的变化:Akt和Her-2发生明显下调,其辅助蛋白Hsp70表达量增多,同时随着17-AAG浓度的增加Bcl-2表达下降,Bax表达上升,说明17-AAG可启动A549和H446细胞的凋亡模式.结论 17-AAG可通过Hsp90信号通路影响A549和H446细胞中Bax和Bcl-2等凋亡蛋白的表达,最终抑制细胞增殖并诱导细胞凋亡.
    목적 관찰17-병희알기-17-거갑양기격이덕매소(17-AAG)대인폐암A549급H446세포조망적영향,초보탐토기가능적작용궤제.방법 실험조세포분별사용농도위50、100、200、400화500 nmol/L적17-AAG진행처리,이불경약물처리조위음성대조,통과MTT법검측12、24、48급72 h4개시간단내17-AAG대A549급H446세포증식적영향.이불경약물처리조위음성대조,통과류식세포술검측농도위100、200화400 nmol/L적17-AAG작용48 h후,대A549급H446세포주기적영향급기촉조망작용.이불경약물처리조위음성대조,통과Western blot법검측농도위100、200화400 nmol/L적17-AAG대A549급H446세포내열휴극단백(Hsp) 90상관단백(Hsp90、Hsp70、Akt화Her-2)화조망상관단백(Bcl-2화Bax)표체적영향.본연구중결과대비류계량자료이(-x)±s표시,채용Origin 8.0통계연건,다조간균수비교채용t검험,조간량량비교용단인소방차분석(One-way ANOVA).결과 17-AAG농도위50～ 500 nmol/L시2개세포주적증식수억제,정현저적농도의뢰성.48 h시,17-AAG대A549급H446세포적반억제농도(IC50)분별위(222±13)화(189 ±7) nmol/L;세포주기실험결과발현,17-AAG가특이성지장A549급H446세포주적세포주기조체우G2/M기;조망실험결과현시,17-AAG가촉진A549급H446세포조망;17-AAG농도위400 nmol/L처리48 h후,A549세포주적세포조망솔위46.3％,H446세포주적세포조망솔위56.9％,여미가약물처리적대조조상비(A549세포주적세포조망솔위11.9％,H446세포주적세포조망솔위6.9％)차이유통계학의의(P＜0.01);경17-AAG처리48 h후,2개세포주중Hsp90신호통로적상관단백균발생료상동추세적변화:Akt화Her-2발생명현하조,기보조단백Hsp70표체량증다,동시수착17-AAG농도적증가Bcl-2표체하강,Bax표체상승,설명17-AAG가계동A549화H446세포적조망모식.결론 17-AAG가통과Hsp90신호통로영향A549화H446세포중Bax화Bcl-2등조망단백적표체,최종억제세포증식병유도세포조망.
    Objective To observe the effect of 17-(allylamino)-17-demethoxygeldanamycin (17-AAG) on the apoptosis of human lung cancer cell lines A549 and H446,and to investigate the potential mechanisms.Methods Proliferation inhibition and apoptosis assays,and the cell cycles were detected by MTT and flow cytometry respectively.Western blot was used to determine the expression level of proteins such as Hsp90,Hsp70,AKt,Her-2,Bcl-2 and Bax.Results After treated with 17-AAG,the proliferation of both A549 and H446 cells was inhibited significantly in a dose-dependent manner; as the concentration of 17-AAG was from 50 to 500 nmol/L,the IC50 values to A549 and H446 cell lines were (222 ± 13) nmol/L and (189 ±7) nmol/L respectively at 48 h.Cell cycle assays showed that 17-AAG was able to arrest cell cycles of A549 and H446 cell lines at the G2/M phase.Apoptosis assay showed that 17-AAGwas capable of inducing apoptosis in A549 and H446 cell lines.After treated with 17-AAG for 48 h,there were significant differences between the 400 nmol/L groups(46.3％ for A549 cell line and 56.9％ for H446 cell line) and the control group (11.9％ for A549 cell line and 6.9％ for H446 cell line,P ＜0.01).Western blot results showed that the relative proteins of the Hsp90 signal pathway in both A549 and H446 cell lines underwent similar changes after 17-AAG treatment:Akt and Her-2 decreased significantly while the expression of Hsp70 increased.Meanwhile,the expression of Bcl-2 decreased but that of Bax increased,indicating that 17-AAG was able to promote apoptosis mode in A549 and H446 cells.Conclusion 17-AAG can regulate the expression level of apoptosis-related proteins such as Bax and Bcl-2 by Hsp90 signaling pathway in A549 and H446 cells,and ultimately inhibit cell proliferation and induce apoptosis.
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