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dc.contributor.authorGuo, X.
dc.contributor.authorEvans, T.R.J.
dc.contributor.authorSomanath, S.
dc.contributor.authorArmesilla, Angel Luis
dc.contributor.authorDarling, John L.
dc.contributor.authorSchatzlein, A.
dc.contributor.authorCassidy, James
dc.contributor.authorWang, Weiguang
dc.date.accessioned2008-06-04T13:16:58Z
dc.date.available2008-06-04T13:16:58Z
dc.date.issued2007
dc.identifier.citationBritish Journal of Cancer, 97(6): 745-754
dc.identifier.issn0007-0920
dc.identifier.pmid17687334
dc.identifier.doi10.1038/sj.bjc.6603930
dc.identifier.urihttp://hdl.handle.net/2436/29507
dc.description.abstractNuclear factor-kappa B (NF-kappaB) is a transcription factor with high transcriptional activity in cancer cells. In this study, we developed a novel enhancer-promoter system, kappaB4-CEA205, in which the basal carcinoembryonic antigen (CEA) promoter sequence (CEA205) was placed downstream of the four tandem-linked NF-kappaB DNA-binding sites (kappaB4). In combination with a kappaB4 enhancer, the transcriptional activity of the CEA promoter was significantly enhanced (three- to eight-fold) in cancer cell lines but not in normal cells. In cancer cell lines, the transcriptional activity of kappaB4-CEA205 was comparable with that of the SV40 promoter. We also constructed vectors in which the thymidine phosphorylase (TP) cDNA was under the control of CEA205, kappaB4, kappaB4-CEA205 and CMV promoters, respectively. TP protein and enzyme activity were detected at comparable levels in kappaB4-CEA205- and CMV-driven TP cDNA-transfected cancer cell lines (H630 and RKO). The kappaB4-TP and CEA205-TP-transfected cell lines, respectively, only demonstrated negligible and low levels of TP protein and enzyme activity. Both CMV- and kappaB4-CEA205-driven TP cDNA transiently transfected cells were 8- to 10-fold sensitised to 5-fluorouracil (5-FU) prodrug, 5'-deoxy-5-fluorouradine (5'-DFUR), in contrast to only 1.5- to 2-fold sensitised by the kappaB4- and CEA205-driven TP cDNA-transfected cells. The bystander killing effect of CMV- and kappaB4-CEA205-driven TP cDNA-transfected cells was comparable. This is the first report that indicates that the NF-kappaB DNA-binding site could be used as a novel cancer-specific enhancer to improve cancer-specific promoter activity in gene-directed enzyme prodrug therapy.
dc.language.isoen
dc.publisherNature Publishing Group
dc.relation.urlhttp://www.nature.com/bjc/journal/v97/n6/abs/6603930a.html
dc.subjectCancer, Colorectal
dc.subjectNF-B
dc.subjectCEA
dc.subjectGDEPT
dc.subjectThymidine Phosphorylase
dc.subjectColorectal cancer
dc.subject.meshBlotting, Western
dc.subject.meshCarcinoembryonic Antigen
dc.subject.meshCell Line, Tumor
dc.subject.meshColonic Neoplasms
dc.subject.meshColorectal Neoplasms
dc.subject.meshCytomegalovirus
dc.subject.meshDNA, Complementary
dc.subject.meshFluorouracil
dc.subject.meshGene Therapy
dc.subject.meshHumans
dc.subject.meshNF-kappa B
dc.subject.meshProdrugs
dc.subject.meshPromoter Regions (Genetics)
dc.subject.meshReverse Transcriptase Polymerase Chain Reaction
dc.subject.meshThymidine Phosphorylase
dc.subject.meshTransfection
dc.titleIn vitro evaluation of cancer-specific NF-kappaB-CEA enhancer-promoter system for 5-fluorouracil prodrug gene therapy in colon cancer cell lines.
dc.typeJournal article
dc.identifier.journalBritish Journal of Cancer
html.description.abstractNuclear factor-kappa B (NF-kappaB) is a transcription factor with high transcriptional activity in cancer cells. In this study, we developed a novel enhancer-promoter system, kappaB4-CEA205, in which the basal carcinoembryonic antigen (CEA) promoter sequence (CEA205) was placed downstream of the four tandem-linked NF-kappaB DNA-binding sites (kappaB4). In combination with a kappaB4 enhancer, the transcriptional activity of the CEA promoter was significantly enhanced (three- to eight-fold) in cancer cell lines but not in normal cells. In cancer cell lines, the transcriptional activity of kappaB4-CEA205 was comparable with that of the SV40 promoter. We also constructed vectors in which the thymidine phosphorylase (TP) cDNA was under the control of CEA205, kappaB4, kappaB4-CEA205 and CMV promoters, respectively. TP protein and enzyme activity were detected at comparable levels in kappaB4-CEA205- and CMV-driven TP cDNA-transfected cancer cell lines (H630 and RKO). The kappaB4-TP and CEA205-TP-transfected cell lines, respectively, only demonstrated negligible and low levels of TP protein and enzyme activity. Both CMV- and kappaB4-CEA205-driven TP cDNA transiently transfected cells were 8- to 10-fold sensitised to 5-fluorouracil (5-FU) prodrug, 5'-deoxy-5-fluorouradine (5'-DFUR), in contrast to only 1.5- to 2-fold sensitised by the kappaB4- and CEA205-driven TP cDNA-transfected cells. The bystander killing effect of CMV- and kappaB4-CEA205-driven TP cDNA-transfected cells was comparable. This is the first report that indicates that the NF-kappaB DNA-binding site could be used as a novel cancer-specific enhancer to improve cancer-specific promoter activity in gene-directed enzyme prodrug therapy.


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