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1、518ActaNutrimentaSinica,Dec.,2006,Vol.28No.6萊菔硫烷對人膀胱癌細胞生長的影響及機制研究單毓娟,吳坤,夏薇(哈爾濱醫(yī)科大學營養(yǎng)與食品衛(wèi)生學教研室,哈爾濱150086)【摘要】目的:研究萊菔硫烷(sulforaphane,SFN)對人膀胱癌細胞生長的影響及作用機制。方法:以人膀胱癌細胞系T24細胞為離體研究對象,通過AO/EB熒光染色觀察SFN誘導T24細胞凋亡核形態(tài)改變;采用流式細胞儀檢測SFN對T24細胞周期的影響;RT-PCR法和Westernblot法檢驗SFN在mRNA和蛋白水平上對TrxR表達的影響。結果:(
2、1)10μmol/L和20μmol/LSFN作用T24細胞24h和48h后,熒光顯微鏡下可見細胞核碎裂和凋亡小體的出現(xiàn)。(2)流式細胞儀檢測可見,SFN能使T24細胞周期阻滯于G0/G1期,20μmol/LSFN作用48h后,在G0/G1峰之前出現(xiàn)凋亡峰。(3)RT-PCR結果:10μmol/LSFN作用T24細胞4h,10h,24h,TrxRmRNA表達有所增加,20μmol萊菔硫烷作用10h,24h后,TrxRmRNA表達明顯增加。(4)Westernblot分析表明,10μmol/LSFN作用T24細胞24h和20μmol/LSFN作用T24細胞8h和2
3、4h后,TrxR蛋白表達有明顯增加。結論:SFN能抑制T24細胞的生長,誘導T24細胞凋亡并使其細胞周期阻滯于G0/G1期,其作用機制與誘導TrxR的轉錄和翻譯水平有關。[營養(yǎng)學報,2006,28(6):518-521]關鍵詞:萊菔硫烷;膀胱癌;硫氧還蛋白還原酶;醌還原酶中圖分類號:R151.41文獻標識碼:A文章編號:0512-7955(2006)06-0518-04EFFECTOFSULFORAPHANEONTHEGROWTHOFHUMANBLADDERCANCERCELLANDITSMECHANISMSHANYu-juan,WUKun,XIAWei(De
4、partmentofNutritionandFoodHygiene,PublicHealthCollege,HarbinMedicalUniversity,Harbin150086,China)【Abstract】Objective:Tostudytheeffectofsulforaphane(SFN)onthegrowthofhumanbladdercancercellanditsmechanisminvitro.Method:MorphologicalcharacteristicsofT24cellnucleusinducedbySFNwereobserv
5、edbyAO/EBfluoresceinstaining.TheeffectofSFNontheT24cellcyclewasdeterminedbyflowcytometry.TheexpressionofTrxRatthetranscriptionalandtranslationallevelsweremeasuredbyRT-PCRandWesternblotmethodsrespectively.Results:(1)Afterthecellsweretreatedwith10μmol/Land20μmol/LSFNfor24hand48h,nucle
6、arfragmentationandapoptoticbodywereseenunderfluorescencemicroscope.(2)SFNcouldblockthecellcycleattheG0/G1phaseshowedbyflowcytometry.Moreover,thetreatmentof20μmol/LSFNfor48hcouldcausetheappearanceofsub-G1beforeG0/G1phase.(3)TheexpressionofTrxRmRNAwereincreasedbythetreatmentof10μmol/L
7、SFNfor4h,10h,24h,comparedwiththecontrolgroup.Furthermore,thetreatmentwithhighdoseSFN(20μmol/L)for10hor24hcouldinducetheexpressionofTrxRmRNAmoresignificantly.(4)TheexpressionofTrxRproteininthe10μmol/LSFNfor24hgroupwasaugmentedcomparedwiththecontrolgroup,andafter收稿日期:2006-05-29基金項目:黑龍
8、江省教育廳科學技術研究項目(No.10