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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.

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Expression of the reporter gene in L02 and BEL-7402 delivered by TAT-LHRH-chitosan/DNA complexes.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; RLU, relative light units.
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f12-ijn-9-2879: Expression of the reporter gene in L02 and BEL-7402 delivered by TAT-LHRH-chitosan/DNA complexes.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; RLU, relative light units.

Mentions: To learn whether the gene delivered by TLC could be expressed in the target cells, both BEL-7402 and L02 cells were transfected with TLCDNPs carrying the pGL3-control plasmid DNA, which encodes luciferase. Figure 12 shows that the luciferase activity in BEL-7402 cells was circa 110 times higher than that in L02 cells. Along with the aforementioned data, it indicated the superior selectivity of TLCDNPs for hepatoma cells over normal liver 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)

Expression of the reporter gene in L02 and BEL-7402 delivered by TAT-LHRH-chitosan/DNA complexes.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; RLU, relative light units.
© Copyright Policy
Related In: Results  -  Collection

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

f12-ijn-9-2879: Expression of the reporter gene in L02 and BEL-7402 delivered by TAT-LHRH-chitosan/DNA complexes.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; RLU, relative light units.
Mentions: To learn whether the gene delivered by TLC could be expressed in the target cells, both BEL-7402 and L02 cells were transfected with TLCDNPs carrying the pGL3-control plasmid DNA, which encodes luciferase. Figure 12 shows that the luciferase activity in BEL-7402 cells was circa 110 times higher than that in L02 cells. Along with the aforementioned data, it indicated the superior selectivity of TLCDNPs for hepatoma cells over normal liver 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