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The Construction of Eukaryotic Expression Vector Containing CYP2B1 Suicide Gene
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- Author:
- No author available
- Journal Title:
- JOURNAL OF CHINESE PHYSICIAN
- Issue:
- 1
- DOI:
- 10.3760/cma.j.issn.1008-1372.2005.01.001
- Key Word:
- CYP2B1/IFF系统;肿瘤;基因治疗
Abstract： 目的构建一种新型的CYP2B1自杀基因表达载体并检测其在肿瘤细胞中的表达.方法根据gene bank中鼠CYP2B1基因的核苷酸序列设计一对引物 ,以 pc3/2B1为模板进行PCR扩增,将PCR CYP2B1基因产物定向插入到pcDNA3.0中,构建CYP2B1基因表达载体pcDNA3.0/CYP2B1;通过酶切、PCR及测序鉴定重组体.采用脂质体介导法将pcDNA3.0/CYP2B1转染三种肿瘤细胞株,RT-PCR检测CYP2B1基因在各细胞株中的表达.结果目的片段均成功插入相应的载体中,CMV启动子调控下的CYP2B1基因在不同的肿瘤细胞株中实现了表达.结论 CMV启动子调控的CYP2B1真核基因表达载体是肿瘤基因治疗中一种新型、高效的治疗载体.
- [1]Marilyn Kozak.Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes[J].1986,44(2).
- [2]Karle P,Salmons B,Renner M,etc.Necrotic, rather than apoptotic, cell death caused by cytochrome P450-activated ifosfamide.[J].2001,8(3).
- [3]M X, Wei,T, Tamiya,R J, Rhee,etc.Diffusible cytotoxic metabolites contribute to the in vitro bystander effect associated with the cyclophosphamide/cytochrome P450 2B1 cancer gene therapy paradigm.[J].1995,1.
- [4]E H, Oldfield,Z, Ram,K W, Culver,etc.Gene therapy for the treatment of brain tumors using intra-tumoral transduction with the thymidine kinase gene and intravenous ganciclovir.[J].1993,4(1).
- [5]M, L?hr,P, Müller,P, Karle,etc.Targeted chemotherapy by intratumour injection of encapsulated cells engineered to produce CYP2B1, an ifosfamide activating cytochrome P450.[J].1998,5(8).
- [6]L, Chen,D J, Waxman.Intratumoral activation and enhanced chemotherapeutic effect of oxazaphosphorines following cytochrome P-450 gene transfer: development of a combined chemotherapy/cancer gene therapy strategy.[J].1995,55.
- [7]G. Brightwell,V. Poirier,E. Cole,etc.Serum-dependent and cell cycle-dependent expression from a cytomegalovirus-based mammalian expression vector[J].1997,194(1).
- [8]J L, Gross,W F, Herblin,B A, Dusak,etc.Effects of modulation of basic fibroblast growth factor on tumor growth in vivo.[J].1993,85.
- [9]Trudeau, C.,Yuan, S.,Galipeau, J.,etc.A Novel Parasite-Derived Suicide Gene for Cancer Gene Therapy with Specificity for Lung Cancer Cells[J].2001,12(13).
- [10]L, Chen,L J, Yu,D J, Waxman.Potentiation of cytochrome P450/cyclophosphamide-based cancer gene therapy by coexpression of the P450 reductase gene.[J].1997,57.
- [11]M, Aghi,T C, Chou,K, Suling,etc.Multimodal cancer treatment mediated by a replicating oncolytic virus that delivers the oxazaphosphorine/rat cytochrome P450 2B1 and ganciclovir/herpes simplex virus thymidine kinase gene therapies.[J].1999,59.
- [12]L, Mohr,N G, Rainov,U G, Mohr,etc.Rabbit cytochrome P450 4B1: A novel prodrug activating gene for pharmacogene therapy of hepatocellular carcinoma.[J].2000,7(7).
- [13]Block A,Freund CT,Woo SL,etc.Gene therapy of metastatic colon carcinoma: regression of multiple hepatic metastases by adenoviral expression of bacterial cytosine deaminase.[J].2000,7(3).
- [14]W F, Anderson.Gene therapy for cancer.[J].1994,5.
- [15]M, Chase,R Y, Chung,E A, Chiocca.An oncolytic viral mutant that delivers the CYP2B1 transgene and augments cyclophosphamide chemotherapy.[J].1998,16.
- [16]M W, Sung,H C, Yeh,S N, Thung,etc.Intratumoral adenovirus-mediated suicide gene transfer for hepatic metastases from colorectal adenocarcinoma: results of a phase I clinical trial.[J].2001,4(3).