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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.

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Cellular uptakes of 9rR-LTVSPWY at different N/P ratios in different cells.(A) Confocal microscopy images of 5-8F and SKOV-3 cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 or 9rR/pEGFP-N1 complexes at the indicated N/P ratios. (B) Percentage of 5-8F cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at the indicated N/P ratios or with a vehicle control (no peptide). Cellular fluorescence was examined by FACS. The data are shown as the means ± SD. All measurements were performed in triplicate. (C) Confocal microscopy images of MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at an N/P ratio of 6. Green fluorescence represents pEGFP-N1 expression. (Scale bar = 20 µm).
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pone-0110632-g003: Cellular uptakes of 9rR-LTVSPWY at different N/P ratios in different cells.(A) Confocal microscopy images of 5-8F and SKOV-3 cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 or 9rR/pEGFP-N1 complexes at the indicated N/P ratios. (B) Percentage of 5-8F cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at the indicated N/P ratios or with a vehicle control (no peptide). Cellular fluorescence was examined by FACS. The data are shown as the means ± SD. All measurements were performed in triplicate. (C) Confocal microscopy images of MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at an N/P ratio of 6. Green fluorescence represents pEGFP-N1 expression. (Scale bar = 20 µm).

Mentions: Because the LTVSPWY peptide has been used as a specific ligand for the cell lines 5–8F and SKOV-3 [23], [27], these two cell lines were used to evaluate the ability of the 9rR-LTVSPWY/pDNA complexes to mediate DNA transfection, while MCF-7 and HeLa cells were used as negative controls. To optimize the peptide/pDNA stoichiometry (N/P ratio) for the best transfection efficiency, the transfection efficiencies of the 9rR-LTVSPWY/pEGFP-N1 complexes at various N/P ratios were measured using GFP as a reporter protein. Confocal microscopy (Figure 3A) showed significant pDNA expression (green fluorescence) in 5–8F and SKOV-3 cells transfected with 9rR-LTVSPWY/pEGFP-N1 at N/P ratios of 6, 9, 12 and 15. N/P = 6 yielded the highest number of cells expressing GFP (Figures 3A, 3B), suggesting that N/P = 6 is the optimal condition for transfection. In contrast, no pDNA expression was observed in MCF-7 and HeLa cells under the same conditions (Figure 3C), demonstrating the cell selectivity of the peptide/pDNA complexes. In contrast, although 9rR encapsulated DNA completely at N/P ratios ≥ 3 [24], there was no obvious GFP expression in the 5–8F cells transfected with the 9rR/pEGFP-N1 complex (Figure 3A and Figure S3), suggesting that the LTVSPWY sequence is essential for the cellular uptake of the 9rR-LTVSPWY/pDNA complexes.


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

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

Cellular uptakes of 9rR-LTVSPWY at different N/P ratios in different cells.(A) Confocal microscopy images of 5-8F and SKOV-3 cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 or 9rR/pEGFP-N1 complexes at the indicated N/P ratios. (B) Percentage of 5-8F cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at the indicated N/P ratios or with a vehicle control (no peptide). Cellular fluorescence was examined by FACS. The data are shown as the means ± SD. All measurements were performed in triplicate. (C) Confocal microscopy images of MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at an N/P ratio of 6. Green fluorescence represents pEGFP-N1 expression. (Scale bar = 20 µm).
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Related In: Results  -  Collection

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pone-0110632-g003: Cellular uptakes of 9rR-LTVSPWY at different N/P ratios in different cells.(A) Confocal microscopy images of 5-8F and SKOV-3 cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 or 9rR/pEGFP-N1 complexes at the indicated N/P ratios. (B) Percentage of 5-8F cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at the indicated N/P ratios or with a vehicle control (no peptide). Cellular fluorescence was examined by FACS. The data are shown as the means ± SD. All measurements were performed in triplicate. (C) Confocal microscopy images of MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes at an N/P ratio of 6. Green fluorescence represents pEGFP-N1 expression. (Scale bar = 20 µm).
Mentions: Because the LTVSPWY peptide has been used as a specific ligand for the cell lines 5–8F and SKOV-3 [23], [27], these two cell lines were used to evaluate the ability of the 9rR-LTVSPWY/pDNA complexes to mediate DNA transfection, while MCF-7 and HeLa cells were used as negative controls. To optimize the peptide/pDNA stoichiometry (N/P ratio) for the best transfection efficiency, the transfection efficiencies of the 9rR-LTVSPWY/pEGFP-N1 complexes at various N/P ratios were measured using GFP as a reporter protein. Confocal microscopy (Figure 3A) showed significant pDNA expression (green fluorescence) in 5–8F and SKOV-3 cells transfected with 9rR-LTVSPWY/pEGFP-N1 at N/P ratios of 6, 9, 12 and 15. N/P = 6 yielded the highest number of cells expressing GFP (Figures 3A, 3B), suggesting that N/P = 6 is the optimal condition for transfection. In contrast, no pDNA expression was observed in MCF-7 and HeLa cells under the same conditions (Figure 3C), demonstrating the cell selectivity of the peptide/pDNA complexes. In contrast, although 9rR encapsulated DNA completely at N/P ratios ≥ 3 [24], there was no obvious GFP expression in the 5–8F cells transfected with the 9rR/pEGFP-N1 complex (Figure 3A and Figure S3), suggesting that the LTVSPWY sequence is essential for the cellular uptake of the 9rR-LTVSPWY/pDNA complexes.

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