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1、2014年第l8卷第11期實用臨床醫(yī)藥雜志JoumalofClinicalMedicineinPractice·1·mytoxinB和myrothecineA抑韋lJ肝癌細胞SMMC-7721增殖的分子機制徐化潔,呂豐偉,程文婷,劉明艷,張瑾,申麗,傅奕(揚州大學(xué)醫(yī)學(xué)院,1.09級臨床本科;2.生物化學(xué)教研室;3.藥學(xué)教研室,江蘇揚州,225001)摘要:目的探討mytoxinB和myrothecineA對肝癌細胞SMMC-7721的增殖抑制作用及分子機制。方法采用MTI"法檢測細胞增殖,流式細胞術(shù)檢測細胞周期變化,Westernblot檢測細胞周
2、期負(fù)調(diào)控蛋白p27蛋白的表達水平及細胞轉(zhuǎn)染psi.p27-469質(zhì)粒(psi—p27),以觀察mytoxinB和myrothecineA是否通過調(diào)節(jié)p27蛋白表達影響SMMC-7721細胞的增殖。結(jié)果mytoxinB和myrothecineA對SMMC-7721細胞增殖皆有較強的抑制作用,且myrothecineA呈現(xiàn)出明顯的劑量依賴性;其增殖抑制作用與S期細胞周期阻滯有關(guān)。進一步研究發(fā)現(xiàn),mytoxinB和myrothecineA皆可導(dǎo)致p27蛋白表達明顯增加;而SMMC-7721細胞在轉(zhuǎn)染psi.p2748h后,p27蛋白的表達明顯減少;同時,
3、myrothecineA所致的細胞增殖抑制完全被阻斷,而mytoxinB所致的細胞增殖抑制部分被逆轉(zhuǎn)。結(jié)論myrothecineA主要通過增加p27蛋白的表達而抑制SMMC-7721細胞增殖,而my'toxinB對SMMC-7721細胞的增殖抑制作用除與上調(diào)p27蛋白表達有關(guān)外,可能還有其他相關(guān)作用機制。關(guān)鍵詞:mytoxinB;myrothecineA;SMMC-7721細胞;增殖中圖分類號:R963文獻標(biāo)志碼:A文章編號:1672-2353(2014)11-001-05DOI:10.7619/jcmp.201411001Molecularmec
4、hanismofmytoxinBandmyrothecineAininhibitingproliferationofSMMC.7721ofhepatocarcinomacellsXUHuajie,LYUFengwei,CHENGWenting。,LIUMingyan,ZHANGJin,SHENLi,F(xiàn)UYi(1.DepartmentofClinicalMedicine;2.DepartmentofBiochemistry;3.DepartmentofPharmacology,SchoolofMedicineofYangzhouUniversity,
5、Yangzhou,Jiangsu,225001)ABSTRACT:ObjectiveToinvestigatethemolecularmechanismofmytoxinBandmyrothecineAininhibitingtheproliferationofSMMC一7721ofhepatocarcinomacells.MethodsMTFassay.flowcytometricanalysis,westernblotandpSi-p27-469plasmid(pSi—p27)celltransfectionexperimentwereused
6、todetecttheproliferationactivity,cellcycledistribution,expressionofnegativecellcycleregula—toryproteinp27andinfluenceofp27oncellproliferation.ResultsTheresultsshowedthatmytoxinBandmyrothecineAexertedpotentanti--proliferativeactivityonSMMC-7721cellsandmyrothecineApresenteddepen
7、dentrelation.Theproliferationinhibitionactivityofthetwocompoundswerebothre—latedtoSphasedelay.FurtherresearchindicatedthatbothmytoxinBandmyrothecineAcouldin—creasetheexpressionofproteinp27obviously.ThepSi-p27celltransfectionexperimentdiscoveredthattheexpressionofproteinp27decr
8、eased觚er48hoursandproliferationinhibitionofmyrothecineAwasblo