Improvement of gene transfer to cervical cancer cell lines using non-viral agents.
Virus-like particles (VLPs) composed of recombinant capsid protein L1 and L2 of human papillomavirus type 16 were conjugated with polylysine (PL) and gene transfer was performed using VLP-PL conjugates to allow the expression of targeted gene. When HeLa cells were incubated with VLP-PL conjugate coupled with plasmid cytomegalovirus beta-galactosidase (pCMVbeta-gal), about 10% of cells were transfected and demonstrated beta-galactosidase activity. Hence chloramphenicol acetyltransferase activity was also expressed significantly in VLP-PL-plasmid simian virus 2 chloramphenicol acetyl transferase (pSV2CAT)-transfected cells, VLP-PL conjugate was tested whether it could transfer a tumor suppressor gene, pCMVp53, to HeLa cells and the exogenously provided p53 gene complexed to VLP-PL conjugate was detected from HeLa cells by polymerase chain reaction (PCR) analysis. Interestingly, additional increase of transfection efficiency was demonstrated in the presence of poloxamer 407 when C-33A cells were transfected with VLP-PL-pCMVbeta-gal complex. The result support the notion that VLP-PL conjugate may be a promising vector to transfer genetic materials into cancer cells and poloxamer 407 can be used for enhancing the transfection efficiency of VLP-PL conjugate.
Cho
C W
CW
Cellular Biology Laboratory, Molecular and Cell Biology Research Division, Korea Research Institute of Bioscience and Biotechnology, P.O. Box 115, Yuseong, Taejon 305-600, South Korea.
Cho
Y S
YS
Kang
B T
BT
Hwang
J S
JS
Park
S N
SN
Yoon
D Y
DY
D002213:Capsid / Q000008:administration & dosage*; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006367:HeLa Cells; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics*; D010957:Plasmids; D011107:Polylysine / Q000008:administration & dosage*; D014162:Transfection
D011107:Polylysine