NDRG2在肿瘤糖代谢中的作用与机制研究

硕士学位论文NDRG2在肿瘤糖代谢中的作用与机制研究李剑英培养类别全日制学位类型专业学位一级学科(专业类)临床医学二级学科(专业)临床检验诊断学研究方向肿瘤代谢指导教师药立波教授培养单位基础部生化教研室二O一三年五月第四军医大学TheFourthMilitaryMedicalUniversity分类号R73UDC616-006密级第四军医大学硕士学位论文独独创创性性声声明明秉承学校严谨的学风与优良的科学道德，本人声明所呈交的论文是我个人在导师指导下进行的研究工作及取得的研究成果。尽我所知，除了文中特别加以标注和致谢的地方外，论文中不包含其他人已经发表或撰写过的研究成果，不包含本人或他人已申请学位或其他用途使用过的成果。与我一同工作的同志对本研究所做的任何贡献均已在论文中作了明确的说明并表示了致谢。申请学位论文与资料若有不实之处，本人承担一切相关责任。论文作者签名：日期：保保护护知知识识产产权权声声明明本人完全了解第四军医大学有关保护知识产权的规定，即：研究生在校攻读学位期间论文工作的知识产权单位为第四军医大学。本人保证毕业离校后，发表论文等使用本论文工作成果时第一署名单位仍然为第四军医大学。学校可以公布论文的全部或部分内容(含电子版，保密内容除外)，可以采用影印，缩印或其他复制手段保存论文；学校有权允许论文被查阅和借阅，并在校园网上提供论文内容的浏览和下载服务。同意学校将论文加入《中国优秀博硕士学位论文全文数据库》和编入《中国知识资源总库》等，并可浏览和下载，同时享受《中国优秀博硕士学位论文全文数据库出版章程》规定的相关权益。论文作者签名：导师签名：日期：第四军医大学硕士学位论文目录缩略语表·······························································································1中文摘要·······························································································5ABSTRACT····························································································7前言······························································································10文献回顾······························································································11一、肿瘤代谢相关综述·······································································11二、NDRG家族相关综述····································································19正文······························································································23第一部分结肠癌HT29细胞NDRG2过表达后物质代谢相关基因表达分析······231材料····························································································232方法····························································································243结果···························································································274讨论····························································································30第二部分NDRG2抑制肿瘤糖酵解代谢表型··············································321材料····························································································322方法····························································································353结果····························································································404讨论····························································································44第三部分NDRG2调控肿瘤糖酵解代谢靶点的筛选与分析····························461材料····························································································462方法····························································································473结果····························································································48第四军医大学硕士学位论文4讨论····························································································55小结······························································································57参考文献······························································································58个人简历和研究成果···············································································68致谢······························································································69第四军医大学硕士学位论文-1-缩略语表缩略词英文全称中文全称AMPKAMP-activatedproteinkinase腺苷酸活化蛋白激酶ATPAdenosineTriphosPhate三磷酸腺苷BpBasepair碱基对BSABovineserumalbumin牛血清白蛋白DEPCDiethylpyrocarbonate二乙基焦碳酸盐DMSODimethylsulfoxide二甲基亚砜dNTPDeoxyribonucleosidetriphosphate脱氧核糖核苷三磷酸2-NBDG(2-[N-(7-nitrobenz-2-oxa-1,3-diaxol-4-yl)amino]-2-deoxyglucose,2-NBDG)2-脱氧葡萄糖ENO1α-enolaseα-希醇化酶F1,6BP1,6-fructose-biphosphate1，6-二磷酸果糖FBSFetalbloodserum胎牛血清G3Pglyceraldehyde3-phosphate3-磷酸甘油醛G6PGlucose-6-phosphate6-磷酸葡萄糖第四军医大学硕士学位论文-2-G6PDGlucose-6-phosphatedehydrogenase葡萄糖-6-磷酸脱氢酶GAPDHGlyceraldehydephosphatedehydrogenase磷酸甘油醛脱氢酶GlUTGlucosetransporter葡萄糖转运蛋白GSHGlutathione谷胱甘肽GSSGOxidizedglutathione氧化型谷胱甘肽HIF-1Hypoxiainduciblefactor-1低氧诱导因子-1HKHexokinase己糖激酶kDaKilodalton千道尔顿kgKilogram千克LLiter升LBLysogenybroth溶菌肉汤LDHLactateDehydrogenase乳酸脱氢酶MMolarconcentration摩尔浓度mAMilliampere毫安mgMilligram毫克第四军医大学硕士学位论文-3-mlMilliliter毫升MTTMethylthiazdyltetrazolium四甲基偶氮唑蓝NADHNicotinamideadeninedinucleotide烟酰胺腺嘌呤二核苷酸NADPHNicotinamideadeninedinucleotidephosphate烟酰胺腺嘌呤二核苷酸磷酸NF-κBNuclearfactorκB核因子κBOXPHOSOxidativePhosphorylation氧化磷酸化PBSPhosphatebufferedsaline磷酸盐缓冲液PDHPyruvatedehydrogenasecomplex丙酮酸脱氢酶复合体PFK-16-phosphofructokinase-16-磷酸果糖激酶-1PETPositronEmissionTomography正电子发射计算机断层仪PFKPhosphofructokinase果糖磷酸激酶PGKPhosphoglyceratekinase1磷酸甘油酸激酶PGMPhosphoglyceratemutase磷酸甘油酸变位酶PKPyruvatekinase丙酮酸激酶PPPPentosephosphatepathway磷酸戊糖途径第四军医大学硕士学位论文-4-PVDFPolyvinylidenefluoride聚偏氟乙烯R5PRibose–5–phosphate5-磷酸核糖RISCRNA-inducedsilencingcomplexRNA诱导的沉默复合物ROSReactiveoxygenspecie活性氧rpmRevolutionsperminute转/分钟SDS-PAGESodiumdodecylsulphate-polyacrylamidegelelectrophoresis十二烷基硫酸钠一聚丙烯酰胺凝胶电泳18F-FDG18F-fluorodeoxyglucose氟代脱氧葡萄糖SGLTSodium/glucosecotransporter钠/葡萄糖协同转运蛋白shRNAShorthairpinRNA短发夹RNAsiRNASmallinterferingRNA小干扰RNATBSTTrisbufferedsalinetween-20Tris吐温缓冲液TEMEDN,N,N,’N’-Tetrameth