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1、A Dissertation Submitted to Huazhong University ofScience and Technology for the Degree ofMaster of SurgeryRECOMBINANT PLASMID PGL3-DF3-DTA FOR THE TREATMENT OF BREAST CANCER: A STUDY OF BALB/C NUDE MOUSE MODELCandidate:Pan ShengMajor:SurgerySupervisor:Prof. Huang Wen GuangHuazhong University of Sci
2、ence & TechnologyWuhan 430074, P. R. ChinaMay, 2007独创性声明本人郑重声明，本学位论文是本人在导师指导下进行的研究工作及取得的研究成果的总结。尽我所知，除文中已经标明引用的内容外，本论文不包含任何其他个人或集体已经发表或撰写过的研究成果。对本文的研究做出贡献的个人和集体，均已在文中以明确方式标明。 本人完全意识到本人将承担本声明引起的一切法律后果。学位论文作者签名：日期：年月日学位论文版权使用授权书本学位论文作者完全了解学校有关保留、使用学位论文的规定，即：学校有权保留并向国家有关部门或机构送交论文的复印件和电子版， 允许论文被查阅和借阅。本人
3、授权华中科技大学可以将本学位论文的全部或部分内容编入有关数据库进行检索，可以采用影印、缩印或扫描等复制手段保存和汇编本学位论文。保密，在 _年解密后适用本授权书。本论文属于不保密。（请在以上方框内打“”）学位论文作者签名：日期：年月日指导教师签名：日期：年月日目录英文缩写一览表 1ABSTRACT 3摘要 5正文 6前言 6材料与方法 8试验结果 15讨论 18结论 25图片 26参考文献 29文献综述 33研究生期间撰写发表的文章 43致谢 44华中科技大学硕士学位论文英文缩写一览表英文缩写英文全称中文全称ADPadenosine diphosphate腺苷二磷酸CEAcarcino-emb
4、ryonic antigen癌胚抗原COXcyclooxygenase环氧化酶COX-1cyclooxygenase-1环氧化酶 -1COX-2cyclooxygenase-2环氧化酶 -2DAB3,3-diaminobenzidine3,3-二氨基联苯胺DCdendritic cell树突状细胞DNAdeoxyribonucleic acid脱氧核糖核酸DTDiphtheria toxin白喉毒素DTADiphtheria toxin A白喉毒素 A 链EDTAethylene diamine tetracetate乙二胺四乙酸EF-2elongation factor-2链延长因子 2EG
5、FRepidermal growth factor receptor表皮生长因子受体ERestrogen receptor雌激素受体GCVganciclovir9-(1,3-二羟 -2-丙氧甲基) 鸟嘌呤GM-CSFgranulocyte-macrophage粒-巨噬细胞集落刺colony-stimulating factor激因子HEHematoxy Eosin stain苏木精伊红染色HER2human epidermal growth人类表皮生长因子受factor receptor2体 2HMFGhuman milk fat globule人乳脂肪球IL2interleukin-2白细胞
6、介素 -2KDkilodalton千道尔顿（质量单位）LBluria-BertaniLB 培养基MDRmultidrug resistance多药耐药1华中科技大学硕士学位论文mRNAmessenger RNA(ribonucleic acid)信使 RNAMMPmatrix metalloproteinases基质金属蛋白酶MUC1mucinl粘蛋白 1MVDmicrovessel density肿瘤微血管密度PBSphosphor-buffered saline磷酸盐缓冲液PC bufferPhosphate-citrate buffer磷酸柠檬酸缓冲液PEpseudomonas exto
7、xin假单胞杆菌外毒素PEGpolyethylene glycol聚乙二醇PGprostaglandin前列腺素PIPropidium Iodide碘化丙啶PKCprotein kinase c蛋白激酶 cPKC-protein kinase c-蛋白激酶 c-PSAPprostate-specific启动子前列腺特异antigen promoter性抗原TIMPtissue inhibitor of metalloproteinase基质金属蛋白酶抑制剂SCIsselective COX-2 inhibitors选择性环氧化酶抑制剂SPFSpecefic Pathogen Free无特定病原
8、体VPFvascular permeability factor血管通透性因子VEGFvascular endothelial growth factor血管内皮生长因子2华中科技大学硕士学位论文Recombinant plasmid PGL3-DF3-DTA for the treatment of breastcancer: a study of Balb/c nude mouse modelAbstractObject Diphtheria toxin A chain (DTA) was introduced as a candidate for cancer cell killing
9、by ribosylating the EF2 translation-initiation factor and inhibiting protein synthesis. Genetic recombination immunotoxin conjugated by the DTA and different vectors can selectively kill cells that express special protein, specially tumor cells. DF3 antigen (also designated MUC1 and episialin) is a
10、member of a family of high molecular weight glycoproteins which are aberrantly overexpressed in over 75% of primary human breast cancers. And it is the relative antigen of human breast cancers which is highly specificity. Expression vector (pGL3-DF3-DTA) containing transcriptional regulation sequenc
11、e of human breast carcinoma DF3/MUC1 promoter and diphtheria toxin A-chain gene（PGL3-DF3-DTA ）inject into the breast cancer xenograft on the nude mouse and observe recombinant plasmid PGL3-DF3-DTA effect of inhibition of human breast cancer growth and cancer tissue apoptosis.Methods 1. To construct the nude mouse modles, the MCF-7 cell line was injected into the subcutaneousnessof the nude mouse ridge. 2. Normal sodium,
