Limits...
TAT-LHRH conjugated low molecular weight chitosan as a gene carrier specific for hepatocellular carcinoma cells.

Liu L, Dong X, Zhu D, Song L, Zhang H, Leng XG - Int J Nanomedicine (2014)

Bottom Line: To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC).In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System.The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People's Republic of China.

ABSTRACT
To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC). TLC/DNA nanoparticles (TLCDNPs) were characterized by agarose gel retardation, atomic force microscopy, and dynamic light scattering analysis. In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System. The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution. The in vitro study demonstrated TLC was highly selective for hepatoma cells and essentially nontoxic.

Show MeSH

Related in: MedlinePlus

Uptake of nanoparticles by BEL-7402 and L02 cells.Notes: (A) Images of BEL-7402 and L02 cells treated under different conditions under High Throughput Screening; (B) mean fluorescence intensity. *P<0.05 versus BEL (TLCDNPs); #P<0.05 versus BEL (TLCDNPs).Abbreviations: BEL, BEL-7402; TLCDNPs, transactivator of transcription – luteinizing hormone-releasing hormone (TAT-LHRH)-chitosan/DNA nanoparticles; Lipo, Lipofectamine® 2000 (Thermo Fisher Scientific, Waltham, MA, USA)/DNA complexes; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein isothiocyanate.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4061174&req=5

f10-ijn-9-2879: Uptake of nanoparticles by BEL-7402 and L02 cells.Notes: (A) Images of BEL-7402 and L02 cells treated under different conditions under High Throughput Screening; (B) mean fluorescence intensity. *P<0.05 versus BEL (TLCDNPs); #P<0.05 versus BEL (TLCDNPs).Abbreviations: BEL, BEL-7402; TLCDNPs, transactivator of transcription – luteinizing hormone-releasing hormone (TAT-LHRH)-chitosan/DNA nanoparticles; Lipo, Lipofectamine® 2000 (Thermo Fisher Scientific, Waltham, MA, USA)/DNA complexes; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein isothiocyanate.

Mentions: To investigate the targeting specificity of TLC for hepatoma cells, both hepatoma cells (BEL-7402) and normal liver cells (L02) were transfected with TLCDNPs or lipoplexes prepared with fluorescein-labeled DNA. As shown in Figure 10, analysis with the GE IN Cell Analyzer 2000 High-Content Cellular Image System (GE Healthcare Bio-Sciences Corp.) demonstrated that the amount of DNA delivered into BEL-7402 cells by TLC was 14 times of that into L02 cells. It also revealed that the transfection efficiency of TCL to hepatoma cells was about 20-fold higher than that of the commercial transfection reagent Lipofectamine 2000, which demonstrated no significant specificity for hepatoma cells.


TAT-LHRH conjugated low molecular weight chitosan as a gene carrier specific for hepatocellular carcinoma cells.

Liu L, Dong X, Zhu D, Song L, Zhang H, Leng XG - Int J Nanomedicine (2014)

Uptake of nanoparticles by BEL-7402 and L02 cells.Notes: (A) Images of BEL-7402 and L02 cells treated under different conditions under High Throughput Screening; (B) mean fluorescence intensity. *P<0.05 versus BEL (TLCDNPs); #P<0.05 versus BEL (TLCDNPs).Abbreviations: BEL, BEL-7402; TLCDNPs, transactivator of transcription – luteinizing hormone-releasing hormone (TAT-LHRH)-chitosan/DNA nanoparticles; Lipo, Lipofectamine® 2000 (Thermo Fisher Scientific, Waltham, MA, USA)/DNA complexes; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein isothiocyanate.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061174&req=5

f10-ijn-9-2879: Uptake of nanoparticles by BEL-7402 and L02 cells.Notes: (A) Images of BEL-7402 and L02 cells treated under different conditions under High Throughput Screening; (B) mean fluorescence intensity. *P<0.05 versus BEL (TLCDNPs); #P<0.05 versus BEL (TLCDNPs).Abbreviations: BEL, BEL-7402; TLCDNPs, transactivator of transcription – luteinizing hormone-releasing hormone (TAT-LHRH)-chitosan/DNA nanoparticles; Lipo, Lipofectamine® 2000 (Thermo Fisher Scientific, Waltham, MA, USA)/DNA complexes; DAPI, 4′,6-diamidino-2-phenylindole; FITC, fluorescein isothiocyanate.
Mentions: To investigate the targeting specificity of TLC for hepatoma cells, both hepatoma cells (BEL-7402) and normal liver cells (L02) were transfected with TLCDNPs or lipoplexes prepared with fluorescein-labeled DNA. As shown in Figure 10, analysis with the GE IN Cell Analyzer 2000 High-Content Cellular Image System (GE Healthcare Bio-Sciences Corp.) demonstrated that the amount of DNA delivered into BEL-7402 cells by TLC was 14 times of that into L02 cells. It also revealed that the transfection efficiency of TCL to hepatoma cells was about 20-fold higher than that of the commercial transfection reagent Lipofectamine 2000, which demonstrated no significant specificity for hepatoma cells.

Bottom Line: To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC).In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System.The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People's Republic of China.

ABSTRACT
To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC). TLC/DNA nanoparticles (TLCDNPs) were characterized by agarose gel retardation, atomic force microscopy, and dynamic light scattering analysis. In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System. The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution. The in vitro study demonstrated TLC was highly selective for hepatoma cells and essentially nontoxic.

Show MeSH
Related in: MedlinePlus