[Study on the preparation of TK-gene nanoparticles and its expression].
OBJECTIVE
To prepare TK-gene nanoparticles and investigate its expression.


METHODS
Biodegradable and biocompatible polymer polylactic-co-glycolic acid (PLGA) was used to prepare recombinant plasmid pEGFP-AFP nanoparticles by double-emulsion evaporation technique. The characteristics of the nanopticicles including morphology, entrapment efficiency was investigated. The expression of TK gene was also investigated by MTT assay, which could determine the dying cells after the addition of gancyclovir (GCV). The enhanced green fluorescent protein (EGFP) expression in human hepatocarcinoma SMMC-7221 cells and human normal parenchymal Chang liver cells were assessed by flow cytometric analysis.


RESULTS
The resulting plasmid-nanoparticles had regular spherical surface and a narrow particle size with a mean diameter of (72 +/- 12) nm, The average entrapment efficiency was 91.25%, the enhanced transfection efficiency and ability protecting plasmid DNA from degraded by nuclease or sonication due to nanoparticles encapsulation.


CONCLUSIONS
DNA-nanoparticles need further study as gene delivery system.
School of Pharmacy, Sichuan University, Chengdu 610041, China. QinHe317@vip.sina.com
Qin
Q
He
Zhi-rong
ZR
Zhang
Ji
J
Liu
Chao-qun
CQ
Xu
D001672:Biocompatible Materials; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D045744:Cell Line, Tumor; D016503:Drug Delivery Systems; D015774:Ganciclovir / Q000494:pharmacology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D018259:Herpesvirus 1, Human / Q000201:enzymology; D006801:Humans; D019344:Lactic Acid; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D036103:Nanotechnology; D010316:Particle Size; D010957:Plasmids; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D011994:Recombinant Proteins / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D000509:alpha-Fetoproteins / Q000235:genetics / Q000378:metabolism
D001672:Biocompatible Materials; D008164:Luminescent Proteins; D011108:Polymers; D011994:Recombinant Proteins; D000509:alpha-Fetoproteins; D049452:Green Fluorescent Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D013937:Thymidine Kinase; D015774:Ganciclovir