代谢组学在肿瘤研究中的应用.ppt

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1、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2012-3-131.1.代谢组学概况代谢组学概况2.2.代谢与肿瘤的关系代谢与肿瘤的关系3.3.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1.1.代谢组学代谢组学1-1 代谢组学的定义代谢组学的定义代谢组学代谢组学(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分析活细胞中代谢物谱变化的研究领域生物标本生物标本:血液、尿液等体液血液、尿液等体液,组织、细胞提取物、细胞培养液组织、细胞提取物、细胞培养液1-2

2、 代谢组学分析技术代谢组学分析技术-NMR-NMRNMR光谱光谱: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和和31P-NMR1-2 代谢组学分析技术代谢组学分析技术-NMR-NMR1-2 代谢组学分析技术代谢组学分析技术-NMR-NMR1-2 代谢组学分析技术代谢组学分析技术-NMR-NMRWarburg,O.On the origin of cancer cells.Science 123,309314(1

3、956)The Warburg Effect1-2 代谢组学分析技术代谢组学分析技术-MS-MSMS:Mass Spectrometry,质谱质谱 基本原理是基本原理是:将样品中各组分电离成离子束将样品中各组分电离成离子束,进入质量分析器聚集进入质量分析器聚集而得到而得到MS图谱，以确定其质量。图谱，以确定其质量。1-2 代谢组学分析技术代谢组学分析技术-MS-MS1-2 代谢组学分析技术代谢组学分析技术-MS-MSMS:Mass Spectrometry,质谱质谱需联合色谱技术对样品进行前期分离需联合色谱技术对样品进行前期分离(1)GC MS联用联用：GC技术是以气体作为流动相的色技术是以气

4、体作为流动相的色 谱法常用于分离挥发性化合物。谱法常用于分离挥发性化合物。(2)LC MS联用联用：LC技技 术是术是 以液以液 体作体作 为流为流 动相动相 的的色色 谱谱 法适用于分离低法适用于分离低 挥发性或非挥发性、热稳定性挥发性或非挥发性、热稳定性 差差的物质。的物质。2.2.代谢与肿瘤的关系代谢与肿瘤的关系 2-1 Tumor glucose metabolic phenotypes:Glucose glycolysis and oxidative phosphorylationWarburg,O.On the origin of cancer cells.Science 123,

5、309314(1956)The Warburg Effect 2-2 Tumor glucose metabolic phenotypes:The pentose phosphate pathway 2-3 Tumor lipid metabolic phenotypes:Simplified overview of tumor lipid metabolismJournal of Nuclear Medicine 2008,Vol.49 No.Suppl_2 43S-63S 2-4 Tumor amino acid metabolic phenotypes:Regulation of gly

6、colysis,glutaminolysis and de novo nucleotide biosynthesis in tumor cells.Current Opinion in Genetics&Development Volume19,Issue1,February2009Cell September5,2008,134,703-707Cancer Cell June2008,13,472-4822.1 P53 2.1 P53 与与肿瘤代谢肿瘤代谢 2-1.p53 in tumor cell metabolism:glycolysisFigure1The three metaboli

7、c fates of glucose in cells and the role for p53 in glucose metabolismnature cell biology VOLUME13|NUMBER3|MARCH2011Bensaad,K.etal.Cell126,107120(2006).2-1 p53 in tumor cell metabolism:glycolysisOnetargetofp53knowntoaffectglucoseglycolysisisTIGAR 2-1 p53 in tumor cell metabolism:glycolysisOnetargeto

8、fp53knowntoaffectglucoseglycolysisisTIGAR 2-1 p53 in tumor cell metabolism:glycolysisOnetargetofp53knowntoaffectglucoseglycolysisisTIGARBensaad,K.etal.Cell126,107120(2006)thefourgenes(pfkfb14)encodingtheenzyme6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase(PFK-2/FBPase-2)2-1 p53 in tumor cell m

9、etabolism:oxidative phosphorylationp53Cell 126,July14,2006Onetargetofp53knowntoaffectglucoseoxidativephosphorylationisSCO2 2-1 p53 in tumor cell metabolism:oxidative phosphorylationOnetargetofp53knowntoaffectglucoseoxidativephosphorylationisSCO2 2-1 p53 in tumor cell metabolism:pentose phosphate pat

10、hwayFigure1The three metabolic fates of glucose in cells and the role for p53 in glucose metabolismnature cell biology VOLUME13|NUMBER3|MARCH2011 2-1 p53 in tumor cell metabolism:pentose phosphate pathway 2-1 p53 in tumor cell metabolism:pentose phosphate pathwayFigure 1.p53 deciency correlates with

11、 increases in PPP flux,glucose consumption and lactate production 2-1 p53 in tumor cell metabolism:pentose phosphate pathwayFigure 2.p53 regulates NADPH levels,lipid accumulation and G6PD activity 2-1 p53 in tumor cell metabolism:pentose phosphate pathwayFigure3p53interactswithG6PDandinhibitsitsacti

12、vityindependentlyoftranscription.2-1 p53 in tumor cell metabolism:pentose phosphate pathwayFigure4p53inhibitstheformationofdimericG6PDholoenzyme.p53regulationofenergymetabolism 2-1 p53 in tumor cell metabolism 2-1 p53 in tumor cell metabolism:glutamine metabolismBiochemistry of glutamine metabolism(

13、main pathways):glutaminase 2-1 p53 in tumor cell metabolism:glutamine metabolism 2-1 p53 in tumor cell metabolism:glutamine metabolismModel for regulation of intracellular ROS levels by GLS2.2.2 HIF-1 2.2 HIF-1 与与肿瘤代谢肿瘤代谢 2-2 HIF-1 in tumor cell metabolismHIF-1:upstream and downstream of cancer meta

14、bolism.CurrOpinGenetDev.2010Feb;20(1):51-6.2-2 HIF-1 in tumor cell metabolismHIF-1:activatingtranscriptionofgenesencodingglucosetransportersandglycolyticenzymeExpressionofgenesencodingglucosetransportersandglycolyticenzymes.Theglycolyticpathwayisshownatleft.Symbolsforgenesencodingtherespectiveenzyme

15、sarecodedbyfontaccordingtothemRNAexpressionpattern(normalizedto18SrRNA)inEScellsculturedundernonhypoxic(N)orhypoxic(H)conditionsfor16hr(lanes16atright)asfollows:(1)(bold)increasedexpressioninhypoxicHif1a+/+cells,lossofinductioninHif1a+/cells,andlossofbasalandinducedexpressioninHif1a/cells;(2)(boldan

16、ditalicized)noeffectofhypoxiaonexpressioninHif1a+/+cellsbutdecreasedexpressioninhypoxicHif1a+/andHif1a/cells;(3)(italicized)noeffectofhypoxiaonexpressioninHif1a+/+cellsbutdecreasedexpressioninhypoxicandnonhypoxicHif1a/cells;(4)(plain)noeffectofhypoxiaorHIF-1deficiencyonexpression.mRNAexpressioninHep

17、3Bcellswasalsoassayed(lanes7,8).Theindicatedgenesencodethefollowingproteins:(GLUT1andGLUT3)glucosetransporter1and3;(HK1andHK2)hexokinase1and2;(GPI)glucosephosphateisomerase;(PFKL)phosphofructokinaseL;(ALDAandALDC)aldolaseAandC;(TPI)triosephosphateisomerase;(GAPDH)glyceraldehyde-3-phosphatedehydrogen

18、ase;(PGK1)phosphoglyceratekinase1;(PGM)phosphoglucomutase;(ENO1)enolase1;(PKM)pyruvatekinaseM;(LDHA)lactatedehydrogenaseA.GLUCOSE(EXT)andGLUCOSE(INT)refertoextracellularandintracellularglucose,respectively,GenesDev.1998Jan15;12(2):149-62.2-2 HIF-1 in tumor cell metabolismHIF-1:up-regulatedtheplasmam

19、embranelactatetransporterMCT4TheplasmamembranelactatetransporterMCT4,butnotMCT1,isup-regulatedbyhypoxiathroughaHIF-1a-dependentmechanism.JBiolChem2006,281:9030-9037.2-2 HIF-1 in tumor cell metabolismPDKPDHHIF-1-mediatedexpressionofpyruvatedehydrogenasekinase:Ametabolicswitchrequiredforcellularadapta

20、tiontohypoxia CELL METABOLISM 3,177185,MARCH2006HIF-1mediatesadaptationtohypoxiabyactivelydownregulatingmitochondrialoxygenconsumptionCELL METABOLISM 3,187197,MARCH2006 2-2 HIF-1 in tumor cell metabolismFigure 1.HIF-1-dependent induction of PDK1 expression in hypoxic cellsFigure6.Coordinateregulatio

21、nofhypoxia-inducedmetabolicswitchesbyHIF-1 2-2 HIF-1 in tumor cell metabolism2.3 mTOR 2.3 mTOR 与与肿瘤代谢肿瘤代谢 2-3 mTOR in tumor cell metabolism Mammalian target of rapamycin up-regulation of pyruvate kinase isoenzyme type M2 is critical for aerobic glycolysis and tumor growth 2-3 mTOR in tumor cell meta

22、bolism:PPP/lipid synthesis Molecular Cell,Volume39,Issue2,171-183,30July2010Highlights:1.mTORC1 stimulates glucose uptake and glycolysis through HIF1 2.mTORC1 stimulates the pentose phosphate pathway and lipid biosynthesis through SREBP1SREBP:sterolregulatoryelement-bindingprotein,SREBPsbelongtotheb

23、asichelix-loop-helixleucinezipper(bHLH-Zip)familyoftranscriptionfactors 2-3 mTOR in tumor cell metabolism:PPP/lipid synthesis J Clin Invest.2002;109(9):11251131 2-3 mTOR in tumor cell metabolism:lipid synthesis mTORC1signalingwillcontributetotumordevelopmentandprogression,thestimulationofribosomebio

24、genesisleadingtoanoverallincreaseinproteinsynthesisislikelytobeamajormechanismbywhichmTORC1promotesanaboliccellgrowthandproliferationintumorcells.2-3 mTOR in tumor cell metabolism J Mol Med(2011)89:221228 2-4 Summary?LDHAMCT4PDHGlutaminaseFASSREBP1/2二氯乙酸DCA丙酮酸脱氢酶临床实验3.3.代谢组学在肿瘤研究中的应代谢组学在肿瘤研究中的应用用 3-

25、1 代谢组学与肿瘤诊断代谢组学与肿瘤诊断 3-1 代谢组学与肿瘤诊断代谢组学与肿瘤诊断 3-1 代谢组学与肿瘤诊断代谢组学与肿瘤诊断 3-1 代谢组学与肿瘤诊断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代谢组学与肿瘤治疗 3-2 代谢组学与肿瘤治疗代谢组学与肿瘤治疗 3-2 代谢组学与肿瘤治疗代谢组学与肿瘤治疗Fig.2.GrowthofmurinemelanomacellsB16F10incultureinarginine-freeRPMI1640mediumwith5%dialysedfetalcalfserumsupplementedwith1mMconcentrationsofarginine(),citrulline()orornithine().Carcinogenesis:metabolic reprogramming Thanks