Folate receptor-mediated intracellular delivery of recombinant caspase-3 for inducing apoptosis.
Recombinant reversed caspase-3 (rev-caspase-3) is a pro-apoptotic gene capable of intracellular autocatalytic processing, which leads to programmed cell death. Folate receptor-specific intracellular delivery of the rev-caspase-3 gene into KB cells over-expressing folate receptors was explored by employing the folate-poly(ethylene glycol)-polyethylenimine (FOL-PEG-PEI) conjugate as a nonviral polymeric carrier. Using luciferase as a reporter gene, the conditions for formulation of DNA/polymer polyplexes were pre-optimized to attain the highest folate receptor-mediated gene transfection efficiency. FOL-PEG-PEI conjugate complexed with rev-caspase-3 plasmid in an optimized condition gave rise to a great increase in expression and activation of exogenous rev-caspase-3 in KB cells when pretreated with doxorubicin. The synthesized conjugate exhibited higher transfection efficiency than other commercially available transfection agents due to a unique mechanism of folate-receptor mediated endocytic gene transfer. The transfected cells showed a significant extent of apoptosis by rev-caspase-3. This study suggests the potential of using folate-receptor-mediated delivery of rev-caspase-3 gene for inducing tumor cell death in a target-specific manner.
Chul Cho
Kyung
K
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, South Korea.
Hoon Jeong
Ji
J
Jung Chung
Hyun
H
Joe
Cheol O
CO
Wan Kim
Sung
S
Gwan Park
Tae
T
D017209:Apoptosis; D002352:Carrier Proteins / Q000096:biosynthesis*; D053148:Caspase 3; D020169:Caspases / Q000235:genetics* / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid / Q000737:chemistry; D018014:Gene Transfer Techniques; D006801:Humans; D007624:KB Cells; D015394:Molecular Structure; D010316:Particle Size; D010957:Plasmids; D011092:Polyethylene Glycols / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D011956:Receptors, Cell Surface / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
D002352:Carrier Proteins; D004337:Drug Carriers; D058973:Folate Receptors, GPI-Anchored; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D005492:Folic Acid; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases