[Constructure of a recombinant adenovirus vector with HSV-TK and it's killer effect on tumor cells in vitro].
High-titer replication-defective recombinant adenovirus expressing HSV-TK gene was constructed. Firstly, shuttle plasmid pAdCMVTK containing HSV-TK gene and CMV promoter was constructed and then recombined with right arm of adenovirus DNA. Secondly, the positive plaques containing recombinant adenovirus were identified and selected out by PCR and Southern blotting after infection into human embryo kidney 293 cells. The titer of recombinant adenovirus AdCMVTK was determined by plaque forming assay and it was as high as 10(12) pfu/ml. Tumor cells were infected with AdCMVTK and then treated with GCV. Cytotoxic effects were assayed with MTT method. HeLa, A549 and LoVo cells infected with AdCMVTK (M. O. I. = 100) became sensitive to the prodrug GCV, with IC50 less than 4 mumol/L. Significant bystander effect was observed. Results here show that the AdCMVTK/GCV system might be potential in the gene therapy for cancer.
Xu
D H
DH
Shanghai Institute of Biochemistry, Chinese Academy of Sciences, Shanghai 200031.
Ge
K
K
Zheng
Z C
ZC
Jian
Q
Q
Liu
X Y
XY
D000256:Adenoviridae / Q000235:genetics*; D002460:Cell Line; D004274:DNA, Recombinant / Q000096:biosynthesis; D004622:Embryo, Mammalian; D015774:Ganciclovir / Q000494:pharmacology; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney / Q000166:cytology; D011401:Promoter Regions, Genetic; D018139:Simplexvirus / Q000737:chemistry; D013937:Thymidine Kinase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects
D004274:DNA, Recombinant; D013937:Thymidine Kinase; D015774:Ganciclovir