Limits...
Selective DNA delivery to tumor cells using an oligoarginine-LTVSPWY peptide.

Gong C, Pan D, Qiu F, Sun P, Zhang YH - PLoS ONE (2014)

Bottom Line: Compared with other non-viral methods such as lipid or polymer-based DNA delivery vectors, peptide-based DNA delivery systems are biocompatible and biodegradable, which leads to lower immunogenicity and lower toxicity.However, peptide-based systems for DNA delivery toward special tumor cells or tissues are still lacking.In this study, we constructed a non-viral 9rR-LTVSPWY peptide-based DNA delivery system and showed that it is able to efficiently and selectively transfect DNA into targeted tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology (HUST), Wuhan, Hubei, China; Key Laboratory of Biomedical Photonics of Ministry of Education, Department of Biomedical Engineering, Huazhong University of Science and Technology, Wuhan, Hubei, China.

ABSTRACT
DNA therapy for cancer requires efficient, selective and safe DNA delivery systems. Compared with other non-viral methods such as lipid or polymer-based DNA delivery vectors, peptide-based DNA delivery systems are biocompatible and biodegradable, which leads to lower immunogenicity and lower toxicity. Moreover, peptide vectors are easier to produce and their compositions easier to control because solid-phase peptide synthesis has been extensively developed. However, peptide-based systems for DNA delivery toward special tumor cells or tissues are still lacking. In this study, we constructed a non-viral 9rR-LTVSPWY peptide-based DNA delivery system and showed that it is able to efficiently and selectively transfect DNA into targeted tumor cells. This work presents a novel strategy for tumor cell-specific DNA delivery and a reference for designing more efficient DNA delivery systems targeted towards various types of cancer.

Show MeSH

Related in: MedlinePlus

Confocal microscopy analysis of the intercellular distributions of 9rR-LTVSPWY/plasmid complexes.Living 5-8F cells were incubated with the 9rR-LTVSPWY/EMA-labeled pEGFP-N1 complex (N/P 6:1) and LysoTracker Green (100 nM) for 30 min at 37°C and post-incubation in RPMI1640 containing 10% FBS for (A) 5 min and (B) 60 min. Rr: Pearson’s coefficient, Red: EMA-labeled pDNA, green: LysoTracker Green, yellow: merge. (Scale bar = 10 µm).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4206439&req=5

pone-0110632-g005: Confocal microscopy analysis of the intercellular distributions of 9rR-LTVSPWY/plasmid complexes.Living 5-8F cells were incubated with the 9rR-LTVSPWY/EMA-labeled pEGFP-N1 complex (N/P 6:1) and LysoTracker Green (100 nM) for 30 min at 37°C and post-incubation in RPMI1640 containing 10% FBS for (A) 5 min and (B) 60 min. Rr: Pearson’s coefficient, Red: EMA-labeled pDNA, green: LysoTracker Green, yellow: merge. (Scale bar = 10 µm).

Mentions: The cellular uptake mechanism of the 9rR-LTVSPWY/pDNA complex was then investigated. 5–8F cells were incubated with EMA-labeled pEGFP-N1 (red) encapsulated with 9rR-LTVSPWY or 9rR. Figure 5A reveals that the EMA-labeled 9rR-LTVSPWY/pDNA complexes (red) were obviously absorbed by the cells in 30 min after incubation and mainly distributed in dots. These puncta co-localized well with the LysoTracker Green probe (green), with a Pearson’s coefficient of 0.794, suggesting that the complexes were mainly located in endosomes and/or lysosomes. In contrast, the 9rR/pDNA complex could not enter the cells at all (Figure S3). When the incubation temperature was decreased from 37°C to 4°C, which typically inhibits endocytosis, the cellular uptake of the 9rR-LTVSPWY/pDNA complex was significantly decreased (Figure 6). These results suggest that the 9rR-LTVSPWY/pDNA complex entered the cells via receptor-mediated endocytosis, consistent with the previously reported cellular uptake mechanism of LTVSPWY [27]. One hour post-incubation, the pDNA (red fluorescence) had escaped from the endosomes (green fluorescence) and was diffusely distributed in the cytosol (Figure 5B), with a resulting decrease in the Pearson’s coefficient (to 0.303). The results showed that pDNA transfected into cells using 9rR-LTVSPWY can escape from endocytic vesicles quickly.


Selective DNA delivery to tumor cells using an oligoarginine-LTVSPWY peptide.

Gong C, Pan D, Qiu F, Sun P, Zhang YH - PLoS ONE (2014)

Confocal microscopy analysis of the intercellular distributions of 9rR-LTVSPWY/plasmid complexes.Living 5-8F cells were incubated with the 9rR-LTVSPWY/EMA-labeled pEGFP-N1 complex (N/P 6:1) and LysoTracker Green (100 nM) for 30 min at 37°C and post-incubation in RPMI1640 containing 10% FBS for (A) 5 min and (B) 60 min. Rr: Pearson’s coefficient, Red: EMA-labeled pDNA, green: LysoTracker Green, yellow: merge. (Scale bar = 10 µm).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4206439&req=5

pone-0110632-g005: Confocal microscopy analysis of the intercellular distributions of 9rR-LTVSPWY/plasmid complexes.Living 5-8F cells were incubated with the 9rR-LTVSPWY/EMA-labeled pEGFP-N1 complex (N/P 6:1) and LysoTracker Green (100 nM) for 30 min at 37°C and post-incubation in RPMI1640 containing 10% FBS for (A) 5 min and (B) 60 min. Rr: Pearson’s coefficient, Red: EMA-labeled pDNA, green: LysoTracker Green, yellow: merge. (Scale bar = 10 µm).
Mentions: The cellular uptake mechanism of the 9rR-LTVSPWY/pDNA complex was then investigated. 5–8F cells were incubated with EMA-labeled pEGFP-N1 (red) encapsulated with 9rR-LTVSPWY or 9rR. Figure 5A reveals that the EMA-labeled 9rR-LTVSPWY/pDNA complexes (red) were obviously absorbed by the cells in 30 min after incubation and mainly distributed in dots. These puncta co-localized well with the LysoTracker Green probe (green), with a Pearson’s coefficient of 0.794, suggesting that the complexes were mainly located in endosomes and/or lysosomes. In contrast, the 9rR/pDNA complex could not enter the cells at all (Figure S3). When the incubation temperature was decreased from 37°C to 4°C, which typically inhibits endocytosis, the cellular uptake of the 9rR-LTVSPWY/pDNA complex was significantly decreased (Figure 6). These results suggest that the 9rR-LTVSPWY/pDNA complex entered the cells via receptor-mediated endocytosis, consistent with the previously reported cellular uptake mechanism of LTVSPWY [27]. One hour post-incubation, the pDNA (red fluorescence) had escaped from the endosomes (green fluorescence) and was diffusely distributed in the cytosol (Figure 5B), with a resulting decrease in the Pearson’s coefficient (to 0.303). The results showed that pDNA transfected into cells using 9rR-LTVSPWY can escape from endocytic vesicles quickly.

Bottom Line: Compared with other non-viral methods such as lipid or polymer-based DNA delivery vectors, peptide-based DNA delivery systems are biocompatible and biodegradable, which leads to lower immunogenicity and lower toxicity.However, peptide-based systems for DNA delivery toward special tumor cells or tissues are still lacking.In this study, we constructed a non-viral 9rR-LTVSPWY peptide-based DNA delivery system and showed that it is able to efficiently and selectively transfect DNA into targeted tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology (HUST), Wuhan, Hubei, China; Key Laboratory of Biomedical Photonics of Ministry of Education, Department of Biomedical Engineering, Huazhong University of Science and Technology, Wuhan, Hubei, China.

ABSTRACT
DNA therapy for cancer requires efficient, selective and safe DNA delivery systems. Compared with other non-viral methods such as lipid or polymer-based DNA delivery vectors, peptide-based DNA delivery systems are biocompatible and biodegradable, which leads to lower immunogenicity and lower toxicity. Moreover, peptide vectors are easier to produce and their compositions easier to control because solid-phase peptide synthesis has been extensively developed. However, peptide-based systems for DNA delivery toward special tumor cells or tissues are still lacking. In this study, we constructed a non-viral 9rR-LTVSPWY peptide-based DNA delivery system and showed that it is able to efficiently and selectively transfect DNA into targeted tumor cells. This work presents a novel strategy for tumor cell-specific DNA delivery and a reference for designing more efficient DNA delivery systems targeted towards various types of cancer.

Show MeSH
Related in: MedlinePlus