中国药理学通报
中國藥理學通報
중국약이학통보
CHINESE PHARMACOLOGICAL BULLETIN
2010年
2期
169-174
,共6页
谢松强%李骞%张亚宏%王建红%赵瑾%王超杰
謝鬆彊%李鶱%張亞宏%王建紅%趙瑾%王超傑
사송강%리건%장아굉%왕건홍%조근%왕초걸
多胺缀合物%多胺转运体%细胞周期%凋亡%Akt%mTOR
多胺綴閤物%多胺轉運體%細胞週期%凋亡%Akt%mTOR
다알철합물%다알전운체%세포주기%조망%Akt%mTOR
polyamine conjugate%polyamine transporter%cell cycle%apoptosis%Akt%mTOR
目的 探讨新型萘酰亚胺-多胺缀合物NNINspm对多胺转运体的识别及诱导肝癌HepG2细胞凋亡的机制.方法 以MTT法检测细胞毒性;流式细胞仪检测细胞周期、凋亡率及线粒体膜电位的变化;高内涵活细胞成像系统检测NNINspm对多胺转运体识别及Akt易位的影响;Western blot检测NNINspm对cytochrome C、14-3-3、Bad、Bcl-xL、mTOR、p70S6K、Cdk4 、p27~(kip1)、Akt、Caspase-3、Caspase-9等蛋白表达的影响.结果 NNINspm具有良好的多胺转运体识别能力及肿瘤细胞靶向性,其通过抑制Akt磷酸化从而引起一系列的信号分子发生改变,如14-3-3蛋白与Bad解离并与Bcl-xL结合,随后引起cytochrome C释放及caspase-9及caspase-3活化并最终诱导细胞凋亡;此外,NNINspm诱导mTOR和p70S6K脱磷酸化,Cdk4下调及 p27~(kip1)上调并最终诱导细胞周期阻滞于G_0/G_1期.结论 NNINspm通过PI_3K/Akt信号途径诱导肝癌HepG2细胞凋亡.
目的 探討新型萘酰亞胺-多胺綴閤物NNINspm對多胺轉運體的識彆及誘導肝癌HepG2細胞凋亡的機製.方法 以MTT法檢測細胞毒性;流式細胞儀檢測細胞週期、凋亡率及線粒體膜電位的變化;高內涵活細胞成像繫統檢測NNINspm對多胺轉運體識彆及Akt易位的影響;Western blot檢測NNINspm對cytochrome C、14-3-3、Bad、Bcl-xL、mTOR、p70S6K、Cdk4 、p27~(kip1)、Akt、Caspase-3、Caspase-9等蛋白錶達的影響.結果 NNINspm具有良好的多胺轉運體識彆能力及腫瘤細胞靶嚮性,其通過抑製Akt燐痠化從而引起一繫列的信號分子髮生改變,如14-3-3蛋白與Bad解離併與Bcl-xL結閤,隨後引起cytochrome C釋放及caspase-9及caspase-3活化併最終誘導細胞凋亡;此外,NNINspm誘導mTOR和p70S6K脫燐痠化,Cdk4下調及 p27~(kip1)上調併最終誘導細胞週期阻滯于G_0/G_1期.結論 NNINspm通過PI_3K/Akt信號途徑誘導肝癌HepG2細胞凋亡.
목적 탐토신형내선아알-다알철합물NNINspm대다알전운체적식별급유도간암HepG2세포조망적궤제.방법 이MTT법검측세포독성;류식세포의검측세포주기、조망솔급선립체막전위적변화;고내함활세포성상계통검측NNINspm대다알전운체식별급Akt역위적영향;Western blot검측NNINspm대cytochrome C、14-3-3、Bad、Bcl-xL、mTOR、p70S6K、Cdk4 、p27~(kip1)、Akt、Caspase-3、Caspase-9등단백표체적영향.결과 NNINspm구유량호적다알전운체식별능력급종류세포파향성,기통과억제Akt린산화종이인기일계렬적신호분자발생개변,여14-3-3단백여Bad해리병여Bcl-xL결합,수후인기cytochrome C석방급caspase-9급caspase-3활화병최종유도세포조망;차외,NNINspm유도mTOR화p70S6K탈린산화,Cdk4하조급 p27~(kip1)상조병최종유도세포주기조체우G_0/G_1기.결론 NNINspm통과PI_3K/Akt신호도경유도간암HepG2세포조망.
Aim To investigate the apoptotic mechanism and polyamine transporter recognition of 3-nitro-naphthalimide norspermine conjugate (NNINspm),a novel naphthalimide-polyamine conjugate, in HepG2 cells.Methods The cytotoxicity of NNINspm was assessed by MTT assay.Cell cycle distribution and apoptosis were measured by flow cytometry.The protein expression of cytochrome C,14-3-3,Bad,Bcl-xL,mTOR,p70S6K,Cdk4,p27~(kip1),Akt,Caspase-3,Caspase-9 was evaluated by Western blot.The translocation of Akt was detected by high content screening (HCS) analysis.Results NNINspm induced HepG2 cells apoptosis via Akt dephosphorylation and then triggered a series of signal events, such as Bad dephosphorylation, dissociation of 14-3-3 and Bad, and then binding to Bcl-xL,which finally resulted in mitochondrial disruption,cytochrome c release and caspase cascade activation.Furthermore,the NNINspm-mediated cell cycle arrest was due to mTOR and p70S6K dephosphorylation,Cdk4 down-regulation and p27~(kip1) up-regulation.Conclusion NNINspm induces HepG2 cell apoptosis via PI_3K/Akt signal pathway.
