木鳖子醇提物诱导黑素瘤B16细胞分化机制及研究.pdfVIP

木鳖子醇提物诱导黑素瘤B16细胞分化机制的研究 赵连梅 韩丽娜 单保恩 河北医科大学第四医院科研中心，河北省石家庄050011 目的：探讨木鳖子醇提物(ethanolextractofcochinchinamomordicaseed，CMSEE)诱导黑素瘤B16细胞分化的作 用机理。方法：CMSEE处理B16细胞后，经瑞氏染色，倒置相差显微镜下观察B16细胞的形态学变化；比色法检测B16细胞 黑素含量。Westernblotting检测经CMSEE和p38、ERK通路抑制剂SB203580和SD98059处理后p-p38、p-ERK表达的变化。结 果：经10μg/ml～40μg/mlCMSEE处理后，B16细胞呈现典型的细胞分化形态，黑色素量含量明显升高 （P＜0.01），；20μg/ml CMSEE处理B16细胞30、60、120、240min后，p-p38蛋白表达水平明显升高(P＜0.01)。p-ERK蛋白表达水平明显降低(P＜0.01)。 总p38和ERK蛋白表达水平不变。p38通路抑制剂SB203580可阻断CMSEE对B16细胞的分化诱导作用，而ERK通路抑制剂可 增强CMSEE对B16细胞的分化诱导作用。结论：p38通路和ERK通路的活性参与了CMSEE对B16细胞的分化诱导作用。 木鳖子醇提物 ;B16;分化；MAPKs StudyforMechanismofEthanolExtractofCochinchinaMomordica SeedonMouseMelanomaB16CellsDifferentiation ZHAOLianmeiHAN LinaSHANBaoen Researchcenter,ThefourthhospitalofHebeimedicaluniversity,Shijiazhuang,HebeiProvince,China 050011 Objective:Tostudythemechanismofethanolextractofcochinchinamomordicaseed(CMSEE) onmousemelanomaB16cellsdifferentiation.Methods:AftertreatedbyCMSEE(10-40μg/ml),morphological changesofB16cellswereobservedbyGiemsastaining.Melanincontentwasassessedbycolorimeter.Western blotwasusedtomeasuretheexpressionofphosphorylationp38、ERKandtotalp38andERK.Result:10,20,40 μg/mlCMSEEcouldinducedifferentiationofB16cellsinvestigatedbymicroscope.Westernblotanalysis revealedthat20μg/mlCMSEEtransientlyelevatedthelevelofp-p38MAPproteinandblocktheexpressionof p-ERKat30min(P＜0.01).whereasCMSEEhasnoeffectonthetotalp38andERKexpression.SB203580,the inhibitorofp38MAP,partlyblockedCMSEE-inducedB16celldifferentiation,butPD98059,theinhibitorof ERKpathway,couldelevateCMSEE-inducedB16celldifferentiation.Conclusion:p38MAPandERKsignal pathwaywereinvolvedinthedifferentiationofB16melanomacells. CMSEE;B16;Differentiation;MAPKsignalpathway L0.5×105/mli ml)，以不j -V¨～ l染色观察 ( )logicalchan ■ hwayinB16ce