2011
DOI: 10.1016/j.canlet.2011.03.016
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Annexin V-targeted enzyme prodrug therapy using cytosine deaminase in combination with 5-fluorocytosine

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Cited by 21 publications
(38 citation statements)
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“…To mimic in vivo EPT, cells were treated with a saturating concentration of fusion protein (100 nM) every 3 days for 2 hours at 37°C in accordance with previous binding stability studies. [18][19][20] Each day, the medium was replaced with a medium containing varying concentrations of prodrug (SM and 5-FC from Fisher Scientific, Waltham, MA and FD from VWR, Radnor, PA) or drug analog (2-FA from Fisher Scientific and 5-FU from Sigma-Aldrich, St. Louis, MO) both containing appropriate concentrations of docetaxel. An Alamar Blue (Invitrogen, Grand Island, NY) assay was preformed every 2 days to measure cell viability.…”
Section: In Vitro Enzyme Prodrug Cytotoxicitymentioning
confidence: 99%
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“…To mimic in vivo EPT, cells were treated with a saturating concentration of fusion protein (100 nM) every 3 days for 2 hours at 37°C in accordance with previous binding stability studies. [18][19][20] Each day, the medium was replaced with a medium containing varying concentrations of prodrug (SM and 5-FC from Fisher Scientific, Waltham, MA and FD from VWR, Radnor, PA) or drug analog (2-FA from Fisher Scientific and 5-FU from Sigma-Aldrich, St. Louis, MO) both containing appropriate concentrations of docetaxel. An Alamar Blue (Invitrogen, Grand Island, NY) assay was preformed every 2 days to measure cell viability.…”
Section: In Vitro Enzyme Prodrug Cytotoxicitymentioning
confidence: 99%
“…Dissociation constants were determined as described previously. [18][19][20] Briefly, cells were fixed with glutaraldehyde, treated with bovine serum albumin, and incubated with varying concentrations (0-20 nM) of biotin-labeled fusion proteins, which after washing were then allowed to react with streptavidin-horseradish peroxidase. Subsequently, binding was quantified with the chromogenic substrate o-phenylenediamine by measuring absorbance at 450 nm.…”
Section: In Vitro Binding Assaysmentioning
confidence: 99%
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“…Cytosine deaminase (CD)/5-fluorocytosine (5-FC), one of the most widely used GEPT systems, is expressed in bacteria like Escherichia coli or yeast like Saccharomyes cerevisiae. In both organisms, the encoded CD catalyzes the conversion of a 5-FC prodrug into a toxic agent, 5-fluorouracil (5-FU), which is able to inhibit cancer cell growth (10,11) by impairing DNA synthesis and promoting apoptosis (12). However, yeast CD (yCD) appears to be far more efficient in the conversion of 5-FC to 5-FU than bacterial CD (bCD) in both in vitro and in vivo models (13).…”
Section: Introductionmentioning
confidence: 99%