代谢组学在肿瘤研究中的应用二

,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,Click to edit Master title style,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,代谢组学在肿瘤研究中的应用二,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,1,H-NMR,、,13,C-NMR,和,31,P-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. On the origin of cancer cells.,Science,123, 309314 (1956),The Warburg Effect,1-2,代谢组学分析技术,-MS,MS: Mass Spectrometry,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: Mass Spectrometry,质谱,需联合色谱技术对样品进行前期分离,( 1 ) GC MS,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 ) LC MS,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,2-1 Tumor glucose metabolic phenotypes:,Glucose glycolysis and oxidative phosphorylation,Warburg, O. On the origin of cancer cells.,Science,123, 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 metabolism,Journal of Nuclear Medicine,2008, Vol. 49 No. Suppl_2 43S-63S,2-4 Tumor amino acid metabolic phenotypes:,Regulation of glycolysis, glutaminolysis and,de novo,nucleotide biosynthesis in tumor cells.,Current Opinion in Genetics & Development,Volume 19, Issue 1, February 2009,Cell,September 5, 2008, 134,703-707,Cancer Cell,June 2008,13,472-482,2. 1 P53,与,肿瘤代谢,2-1. p53 in tumor cell metabolism: glycolysis,Figure 1,The three metabolic fates of glucose in cells and the role for p53 in glucose metabolism,nature cell biology,VOLUME 13 | NUMBER 3 | MARCH 2011,Bensaad, K. et al. Cell 126, 107120 (2006).,2-1 p53 in tumor cell metabolism: glycolysis,One target of p53 known to affect glucose glycolysis is TIGAR,2-1 p53 in tumor cell metabolism: glycolysis,One target of p53 known to affect glucose glycolysis is TIGAR,2-1 p53 in tumor cell metabolism: glycolysis,One target of p53 known to affect glucose glycolysis is TIGAR,Bensaad, K. et al. Cell 126, 107120 (2006),the four genes (pfkfb14) encoding the enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase-2),2-1 p53 in tumor cell metabolism:,oxidative phosphorylation,p53,Cell,126, July 14, 2006,One target of p53 known to affect glucose oxidative phosphorylation is SCO2,2-1 p53 in tumor cell metabolism:,oxidative phosphorylation,One target of p53 known to affect glucose oxidative phosphorylation is SCO2,2-1 p53 in tumor cell metabolism:,pentose phosphate pathway,Figure 1,The three metabolic fates of glucose in cells and the role for p53 in glucose metabolism,nature cell biology,VOLUME 13 | NUMBER 3 | MARCH 2011,2-1 p53 in tumor cell metabolism:,pentose phosphate pathway,2-1 p53 in tumor cell metabolism:,pentose phosphate pathway,Figure 1 . p53 deciency correlates with increases in PPP flux, glucose consumption and lactate production,2-1 p53 in tumor cell metabolism:,pentose phosphate pathway,Figure 2. p53 regulates NADPH levels, lipid accumulation and G6PD activity,2-1 p53 in tumor cell metabolism:,pentose phosphate pathway,Figure 3 p53 interacts with G6PD and inhibits its activity independently of transcription.,2-1 p53 in tumor cell metabolism:,pentose phosphate pathway,Figure 4 p53 inhibits the formation of dimeric G6PD holoenzyme.,p53 regulation of energy metabolism,2-1 p53 in tumor cell metabolism,2-1 p53 in tumor cell metabolism:,glutamine metabolism,Biochemistry of glutamine metabolism (main pathways):,glutaminase,2-1 p53 in tumor cell metabolism:,glutamine metabolism,2-1 p53 in tumor cell metabolism:,glutamine metabolism,Model for regulation of intracellular ROS levels by GLS2.,2. 2 HIF-1,与,肿瘤代谢,2-2 HIF-1 in tumor cell metabolism,HIF-1: upstream and downstream of cancer metabolism.,Curr Opin Genet Dev. 2010 Feb;20(1):51-6.,2-2 HIF-1 in tumor cell metabolism,HIF-1,:,activating transcription of genes encoding glucose transporters and glycolytic enzyme,Expression of genes encoding glucose transporters and glycolytic enzymes. The glycolytic pathway is shown at,left.,Symbols for genes encoding the respective enzymes are coded by font according to the mRNA expression pattern (normalized to 18S rRNA) in ES cells cultured under nonhypoxic (N) or hypoxic (H) conditions for 16 hr (lanes,16,at,right,) as follows: (1) (bold) increased expression in hypoxic,Hif1a,+/+,cells, loss of induction in,Hif1a,+/,cells, and loss of basal and induced expression in,Hif1a,/,cells; (2) (bold and italicized) no effect of hypoxia on expression in,Hif1a,+/+,cells but decreased expression in hypoxic,Hif1a,+/,and,Hif1a,/,cells; (3) (italicized) no effect of hypoxia on expression in,Hif1a,+/+,cells but decreased expression in hypoxic and nonhypoxic,Hif1a,/,cells; (4) (plain) no effect of hypoxia or HIF-1,deficiency on expression. mRNA expression in Hep3B cells was also assayed (lanes,7,8,). The indicated genes encode the following proteins: (,GLUT1,and,GLUT3,) glucose transporter 1 and 3; (,HK1,and,HK2,) hexokinase 1 and 2; (GPI) glucosephosphate isomerase; (,PFKL,) phosphofructokinase L; (,ALDA,and,ALDC,) aldolase A and C; (,TPI,) triosephosphate isomerase; (,GAPDH,) glyceraldehyde-3-phosphate dehydrogenase; (,PGK1,) phosphoglycerate kinase 1; (PGM) phosphoglucomutase; (,ENO1,) enolase 1; (,PKM,) pyruvate kinase M; (,LDHA,) lactate dehydrogenase A. GLUCOSE (EXT) and GLUCOSE (INT) refer to extracellular and intracellular glucose, respectively,Genes Dev. 1998 Jan 15;12(2):149-62.,2-2 HIF-1 in tumor cell metabolism,HIF-1,:,up-regulated the plasma membrane lactate transporter MCT4,The plasma membrane lactate transporter MCT4, but not MCT1, is up-regulated by hypoxia through a HIF-1a-dependent mechanism.,J Biol Chem 2006, 281:9030-9037.,2-2 HIF-1 in tumor cell metabolism,PDK,PDH,HIF-1-mediated expression of pyruvate dehydrogenase kinase: A metabolic switch required for cellular adaptation to hypoxia,CELL METABOLISM,3, 177185,MARCH 2006,HIF-1mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption,CELL METABOLISM,3, 187197,MARCH 2006,2-2 HIF-1 in tumor cell metabolism,Figure 1. HIF-1-dependent induction of PDK1 expression in hypoxic cells,Figure 6. Coordinate regulation of hypoxia-induced metabolic switches by HIF-1,2-2 HIF-1 in tumor cell metabolism,2. 3 mTOR,与,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 metabolism: PPP/lipid synthesis,Molecular Cell,Volume 39, Issue 2,171-183, 30 July 2010,Highlights:,1. mTORC1 stimulates glucose uptake and glycolysis through HIF1,2. mTORC1 stimulates the pentose phosphate pathway and lipid biosynthesis through SREBP1,SREBP: sterol regulatory element-binding protein, SREBPs belong to the basic helix-loop-helixleucine zipper (bHLH-Zip) family of transcription factors,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,mTORC1 signaling will contribute to tumor development and progression, the stimulation of ribosome biogenesis leading to an overall increase in protein synthesis is likely to be a major mechanism by which mTORC1 promotes anabolic cell growth and proliferation in tumor cells.,2-3 mTOR in tumor cell metabolism,J Mol Med (2011) 89:221228,2-4,Summary,?,LDHA,MCT4,PDH,Glutaminase,FAS,SREBP1/2,二氯乙酸,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-glucose molar ratio for each patient sle.,a,normal tissue (NT);,b,cirrhotic tissue (CIR);,c,hepatocellular carcinoma (HCC);,d,liver metastasis from colorectal carcinoma (MET-CRC).,3-2,代谢组学与肿瘤治疗,3-2,代谢组学与肿瘤治疗,3-2,代谢组学与肿瘤治疗,Fig. 2. Growth ofmurinemelanoma cells B16F10 in culture in arginine-free,RPMI 1640 medium with 5% dialysed fetal calf serum supplemented with,1mM concentrations of arginine (), citrulline () or ornithine ().,Carcinogenesis: metabolic reprogramming,