代谢组学在肿瘤研究中的应用

代谢组学在肿瘤研究中的应用,2012-3-13,代谢组学概况代谢与肿瘤的关系代谢组学在肿瘤研究中的应用,1.代谢组学,1-1代谢组学的定义,代谢组学(metabonomics):某一生物或细胞所有低分子质量代谢产物进行定性和定量检测,分析活细胞中代谢物谱变化的研究领域,生物标本:血液、尿液等体液,组织、细胞提取物、细胞培养液,1-2代谢组学分析技术-NMR,NMR光谱:NMR技术是最早被用于代谢组学研究的技术之一其利用原子核在磁场中的能量变化来获得相关信息。目前常用的有1H-NMR、13C-NMR和31P-NMR,1-2代谢组学分析技术-NMR,1-2代谢组学分析技术-NMR,1-2代谢组学分析技术-NMR,Warburg,O.Ontheoriginofcancercells.Science123,309314(1956),TheWarburgEffect,1-2代谢组学分析技术-MS,MS:MassSpectrometry,质谱基本原理是:将样品中各组分电离成离子束,进入质量分析器聚集而得到MS图谱，以确定其质量。,1-2代谢组学分析技术-MS,1-2代谢组学分析技术-MS,MS:MassSpectrometry,质谱需联合色谱技术对样品进行前期分离(1)GCMS联用：GC技术是以气体作为流动相的色谱法常用于分离挥发性化合物。(2)LCMS联用：LC技术是以液体作为流动相的色谱法适用于分离低挥发性或非挥发性、热稳定性差的物质。,2.代谢与肿瘤的关系,2-1Tumorglucosemetabolicphenotypes:Glucoseglycolysisandoxidativephosphorylation,Warburg,O.Ontheoriginofcancercells.Science123,309314(1956),TheWarburgEffect,2-2Tumorglucosemetabolicphenotypes:,Thepentosephosphatepathway,2-3Tumorlipidmetabolicphenotypes:,Simplifiedoverviewoftumorlipidmetabolism,JournalofNuclearMedicine2008,Vol.49No.Suppl_243S-63S,2-4Tumoraminoacidmetabolicphenotypes:,Regulationofglycolysis,glutaminolysisanddenovonucleotidebiosynthesisintumorcells.,CurrentOpinioninGenetics20(1):51-6.,2-2HIF-1intumorcellmetabolism,HIF-1:activatingtranscriptionofgenesencodingglucosetransportersandglycolyticenzyme,Expressionofgenesencodingglucosetransportersandglycolyticenzymes.Theglycolyticpathwayisshownatleft.SymbolsforgenesencodingtherespectiveenzymesarecodedbyfontaccordingtothemRNAexpressionpattern(normalizedto18SrRNA)inEScellsculturedundernonhypoxic(N)orhypoxic(H)conditionsfor16hr(lanes16atright)asfollows:(1)(bold)increasedexpressioninhypoxicHif1a+/+cells,lossofinductioninHif1a+/cells,andlossofbasalandinducedexpressioninHif1a/cells;(2)(boldanditalicized)noeffectofhypoxiaonexpressioninHif1a+/+cellsbutdecreasedexpressioninhypoxicHif1a+/andHif1a/cells;(3)(italicized)noeffectofhypoxiaonexpressioninHif1a+/+cellsbutdecreasedexpressioninhypoxicandnonhypoxicHif1a/cells;(4)(plain)noeffectofhypoxiaorHIF-1deficiencyonexpression.mRNAexpressioninHep3Bcellswasalsoassayed(lanes7,8).Theindicatedgenesencodethefollowingproteins:(GLUT1andGLUT3)glucosetransporter1and3;(HK1andHK2)hexokinase1and2;(GPI)glucosephosphateisomerase;(PFKL)phosphofructokinaseL;(ALDAandALDC)aldolaseAandC;(TPI)triosephosphateisomerase;(GAPDH)glyceraldehyde-3-phosphatedehydrogenase;(PGK1)phosphoglyceratekinase1;(PGM)phosphoglucomutase;(ENO1)enolase1;(PKM)pyruvatekinaseM;(LDHA)lactatedehydrogenaseA.GLUCOSE(EXT)andGLUCOSE(INT)refertoextracellularandintracellularglucose,respectively,GenesDev.1998Jan15;12(2):149-62.,2-2HIF-1intumorcellmetabolism,HIF-1:up-regulatedtheplasmamembranelactatetransporterMCT4,TheplasmamembranelactatetransporterMCT4,butnotMCT1,isup-regulatedbyhypoxiathroughaHIF-1a-dependentmechanism.JBiolChem2006,281:9030-9037.,2-2HIF-1intumorcellmetabolism,PDKPDH,HIF-1-mediatedexpressionofpyruvatedehydrogenasekinase:AmetabolicswitchrequiredforcellularadaptationtohypoxiaCELLMETABOLISM3,177185,MARCH2006,HIF-1mediatesadaptationtohypoxiabyactivelydownregulatingmitochondrialoxygenconsumption,CELLMETABOLISM3,187197,MARCH2006,2-2HIF-1intumorcellmetabolism,Figure1.HIF-1-dependentinductionofPDK1expressioninhypoxiccells,Figure6.Coordinateregulationofhypoxia-inducedmetabolicswitchesbyHIF-1,2-2HIF-1intumorcellmetabolism,2.3mTOR与肿瘤代谢,2-3mTORintumorcellmetabolism,Mammaliantargetofrapamycinup-regulationofpyruvatekinaseisoenzymetypeM2iscriticalforaerobicglycolysisandtumorgrowth,/cgi/doi/10.1073/pnas.1014769108,2-3mTORintumorcellmetabolism:PPP/lipidsynthesis,MolecularCell,Volume39,Issue2,171-183,30July2010,Highlights:1.mTORC1stimulatesglucoseuptakeandglycolysisthroughHIF12.mTORC1stimulatesthepentosephosphatepathwayandlipidbiosynthesisthroughSREBP1,SREBP:sterolregulatoryelement-bindingprotein,SREBPsbelongtothebasichelix-loop-helixleucinezipper(bHLH-Zip)familyoftranscriptionfactors,2-3mTORintumorcellmetabolism:PPP/lipidsynthesis,JClinInvest.2002;109(9):11251131,2-3mTORintumorcellmetabolism:lipidsynthesis,mTORC1signalingwillcontributetotumordevelopmentandprogression,thestimulationofribosomebiogenesisleadingtoanoverallincreaseinproteinsynthesisislikelytobeamajormechanismbywhichmTORC1promotesanaboliccellgrowthandproliferationintumorcells.,2-3mTORintumorcellmetabolism,JMolMed(2011)89:221228,2-4Summary,?LDHAMCT4PDHGlutaminaseFASSREBP1/2,二氯乙酸DCA丙酮酸脱氢酶临床实验,3.代谢组学在肿瘤研究中的应用,3-1代谢组学与肿瘤诊断,3-1代谢组学与肿瘤诊断,3-1代谢组学与肿瘤诊断,3-1代谢组学与肿瘤诊断,Fig.6Lactate-glucosemolarratioforeachpatientsample.,anormaltissue(NT);bcirrhotictissue(CIR);chepatocellularcarcinoma(HCC);dlivermetastasisfromcolorectalcarcinoma(MET-CRC).,3-2代谢组学与肿瘤治疗,3-2代谢组学与肿