Calcium phosphate nanoparticles as a novel nonviral vector for efficient transfection of DNA in cancer gene therapy.
To explore an efficient gene vector in cancer gene therapy, a novel nonviral vector calcium phosphate nanoparticle (CPNP) was developed. Transmission electromicroscopy and Zeta potential demonstrated that CPNP was 23.5-34.5 nm diameters and had +16.8 mV positive surface charges. The analysis of the CPNPDNA complex showed CPNP could transfer foreign DNA into targeted cells with high transfection efficiency, as well as its DNA-binding property and protection of DNA from degradation. Furthermore, the CPNP-DNA complex had no obvious cytotoxicity for CNE-2 cells, while the liposome-DNA complex had certain cytotoxicity. When the CPNP combined with suicide genes yCDglyTK for nasopharyngeal carcinoma (NPC) therapy in vitro, just 24.76% of cells survived when the wild-type CNE-2 cells were treated with the CPNP-yCDglyTK complex plus the prodrug, 5-FC (200 mg/mL). Otherwise, the expression of yCDglyTK was detected in implanted CNE-2 tumors by reverse transcription-polymerase chain reaction (RT-PCR) analysis when the CNE-2 tumor was treated with an intratumoral injection of the CPNPyCDglyTK complex. Our results showed that the CPNP might be a potential vector for gene therapy.
Liu
Ting
T
Xiangya Hospital, Central South University, Changsha 410078, Hunan Province, People's Republic of China.
Tang
Aifa
A
Zhang
Guiying
G
Chen
Yuxiang
Y
Zhang
Junyi
J
Peng
Shusong
S
Cai
Zhiming
Z
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D002130:Calcium Phosphates / Q000737:chemistry*; D002277:Carcinoma / Q000628:therapy; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D005437:Flucytosine / Q000494:pharmacology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000737:chemistry; D008081:Liposomes / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D049329:Nanostructures / Q000737:chemistry*; D009303:Nasopharyngeal Neoplasms / Q000628:therapy; D010316:Particle Size; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D014162:Transfection
D000963:Antimetabolites; D002130:Calcium Phosphates; D017931:DNA Primers; D008081:Liposomes; D013778:Tetrazolium Salts; D013844:Thiazoles; D049452:Green Fluorescent Proteins; D012313:RNA; D004247:DNA; C020243:calcium phosphate; D005437:Flucytosine; C022616:thiazolyl blue