水飞蓟素可通过MAPK信号转导通路促进肺腺癌A549细胞凋亡.docx

水飞蓟素可通过MAPK信号转导通路促进肺腺癌A549细胞凋亡
【摘要】目的：探讨水飞蓟素(SM)对人肺腺癌细胞(A549)的抗癌作用及其分子机制.方法：采用倒置显微镜和电子显微镜等形态学检测、四甲基偶氮唑蓝(MTT)比色法、流式细胞仪(FCM)技术结合PI及AnnexinV双标记染色以及MAPK的磷酸化活性检测方法.结果：SM对人肺腺癌A549细胞的增殖有抑制作用；SM作用A549细胞48h后，随着浓度的增加，倒置显微镜下可见细胞数目减少，有些细胞变小、变圆；透射电镜观察发现，随着SM作用浓度的增加，A549细胞中逐步出现增多的凋亡细胞，凋亡细胞表现出典型的超微结构特征；流式细胞仪检测的结果发现，随着药物作用时间的延长，A549细胞的G1期细胞比例增多，S期细胞明显减少，G2期细胞略有减少，并出现一明显的凋亡峰.SM可以抑制A549细胞中ERK的活性，并在一定程度上提高p38和JNK的活性.结论：SM可以在体外抑制人肺腺癌细胞A549的增殖作用，并诱导其凋亡.SM可能通过抑制A549细胞中ERK的活性，并提高p38和JNK的活性发挥其诱导作用.【关键词】水飞蓟素；肺肿瘤；腺癌；细胞系；凋亡；MAPK信号通路【Abstract】AIM:TostudytheanticancereffectandthemolecularmechanismofsilymarinonhumanlungadenocarcinomacelllineA549.METHODS:Themorphologicalcharactersofapoptosisinducedbysilymarinwerestudiedandanalyzedbyinvertedmicroscopy,electronmicroscopy,methylthiazolyltetrazolium(MTT)colorimetricassay,combinationofflowcytometry(FCM)andAnnexinV/PropidiumIodidedoublestaining,anddetectionofphosphorylationactivityofMAPK.RESULTS:SilymarinhadasignificantinhibitoryeffectontheproliferationofhumanlungadenocarcinomacelllineA549;treatedwithsilymarinfor48h,someofthecellsbecamesmallerandlookedmoreroundthanbefore;ThenumberofA549cellsshowingapoptosisincreasedwithelevationoftheconcentrationofthesilymarin,andthesecellsappearedtypicalultrastructuralfeatures;theresultsofFCMindicatedthattheproportionofA549cellsinG1phaseincreased,whilethenumberofcellsinSphasedecreasedsignificantlyandthoseinG2phasereducedslightlybutpresentedamarkedapoptoticpeak.SilymarinhadanabilityofinhibitingtheactivityofERKandincreasingtheactivitiesofp38andJNKtosomedegreeinA549cells.CONCLUSION:SilymarincaninhibittheproliferationofhumanlungadenocarcinomacelllineA549invitroandinduceitsapoptosispossiblythroughinhibitingtheactivityofERKandincreasingtheactivitiesofp38andJNKinA549cells.【Keywords】silymarin;lungneoplasms;adenocarcinoma;cellline;apoptosis;MAPKsignalpathway0引言水飞蓟素是从植物水飞蓟中提取的一种黄酮类抗氧化剂，具有抗脂质过氧化、清除自由基、抑制5脂氧合酶、维持细胞膜稳定性、促进肝细胞再生及降血脂等作用，具有较强的肝脏保护作用，常用来治疗肝纤维化、乙醇相关性肝病等［1］.近年来的研究发现，SM可以通过诱导肿瘤细胞周期阻滞和凋亡、抑制新生血管生成等机制，抑制如前列腺癌、皮肤癌、结肠癌、白血病等