期刊
CANCER GENE THERAPY
卷 8, 期 6, 页码 397-404出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/sj.cgt.7700310
关键词
heat shock promoter; p53 mutant; HSV-TK; E. coli CD; adenoviral vector; cytopathic effect
类别
资金
- NCI NIH HHS [CA44550, CA44704, CA48000, CA42857] Funding Source: Medline
Tumor cells that express a fusion gene of Escherichia coli cytosine deaminase (CD) and herpes simplex virus type I thymidine kinase (TK) sequences activate and are subsequently killed by the nontoxic prodrugs 5-fluorocytosine and ganciclovir. We have previously developed a recombinant adenovirus containing the CD - TK fusion gene controlled by the human inducible heat shock protein 70 promoter so that heat at 41 degreesC for I hour induces therapeutic gene expression. This adenovirus effectively transduces heat-inducible expression of the CD-TK gene into human prostate carcinoma cells. However, because a limited number of cells in a tumor can actually be infected, we created a replicating adenoviral vector to increase CD-TK gene expression. This vector is a replication-competent, E1B-attenuated adenoviral vector containing the hsp70 promoter-driven CD-TK gene (Ad.E1A(+)HS-CDTK). When human prostate adenocarcinoma DU-145 cells (mutant p53) were infected with the virus at a multiplicity of infection (MOI) of I or 10, the viral replication was detected within 2 days at both MOIs. Similar results were observed in human colorectal carcinoma CX-I cells. When DU-145 cells were infected with the virus at an MOI of 10, incubated for 24 hours, heated at 41 degreesC for 4 hours and then harvested 20 hours later, Western blot analysis demonstrated that this virus successfully produced viral EIA proteins and heat shock stimulated the CD-TK gene expression by 12.3-fold. In addition, Ad.E1A(+)HS-CDTK effectively suppressed cell proliferation by viral cytopathic, effect). Unlike with a replication-incompetent virus (Ad.HS-CDTK), the cytopathic effect of the virus and cytotoxicity in the presence of the prodrugs were still observed even at low MOI (MOI=1.0).
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据