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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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Comparison of the cellular uptakes of 9rR-LTVSPWY in different cells.(A) Percentage of 5-8F, SKOV-3, MCF-7 and HeLa cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes (N/P 6:1, black bars) or a vehicle control (blank, white bars). (B) Quantitative evaluation of luciferase expression in 5-8F, SKOV-3, MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pGL3 complex (N/P 6:1, black bars) or a vehicle control (blank, white bars). The data are shown as the means ± SD. All measurements were performed in triplicate. The “***” symbol indicates a significant difference between the two groups based on three independent experiments (p<0.001). The “#” symbol indicates no significant difference between the two groups based on three independent experiments (p>0.05).
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pone-0110632-g004: Comparison of the cellular uptakes of 9rR-LTVSPWY in different cells.(A) Percentage of 5-8F, SKOV-3, MCF-7 and HeLa cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes (N/P 6:1, black bars) or a vehicle control (blank, white bars). (B) Quantitative evaluation of luciferase expression in 5-8F, SKOV-3, MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pGL3 complex (N/P 6:1, black bars) or a vehicle control (blank, white bars). The data are shown as the means ± SD. All measurements were performed in triplicate. The “***” symbol indicates a significant difference between the two groups based on three independent experiments (p<0.001). The “#” symbol indicates no significant difference between the two groups based on three independent experiments (p>0.05).

Mentions: Because the confocal imaging and FACS results suggested that N/P = 6 results in the highest transfection efficiency, we conducted further experiments with the peptide/pDNA complexes prepared at an N/P ratio of 6. To quantify the specificity and efficiency of transfection in various types of cells, we performed quantitative analysis using flow cytometry. Figure 4A showed that the percentages of 5–8F, SKOV-3, MCF-7 and HeLa cells expressing GFP were approximately 30.33, 8.23, 1.04 and 1.05, respectively, implying that the plasmids were expressed in 5–8F and SKOV-3 cells, with significantly different expression levels in the experimental groups and the blank group, but not to MCF-7 or HeLa, which exhibited no significant difference between the experimental groups and the blank group. Next, we performed luciferase expression assays using the four cell lines. Figure 4B indicates that 9rR-LTVSPWY delivered the plasmid into 5–8F and SKOV-3 cells but not MCF-7 or HeLa cells, verifying the excellent selectivity of this method.


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

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

Comparison of the cellular uptakes of 9rR-LTVSPWY in different cells.(A) Percentage of 5-8F, SKOV-3, MCF-7 and HeLa cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes (N/P 6:1, black bars) or a vehicle control (blank, white bars). (B) Quantitative evaluation of luciferase expression in 5-8F, SKOV-3, MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pGL3 complex (N/P 6:1, black bars) or a vehicle control (blank, white bars). The data are shown as the means ± SD. All measurements were performed in triplicate. The “***” symbol indicates a significant difference between the two groups based on three independent experiments (p<0.001). The “#” symbol indicates no significant difference between the two groups based on three independent experiments (p>0.05).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0110632-g004: Comparison of the cellular uptakes of 9rR-LTVSPWY in different cells.(A) Percentage of 5-8F, SKOV-3, MCF-7 and HeLa cells expressing GFP at 48 h after transfection with the 9rR-LTVSPWY/pEGFP-N1 complexes (N/P 6:1, black bars) or a vehicle control (blank, white bars). (B) Quantitative evaluation of luciferase expression in 5-8F, SKOV-3, MCF-7 and HeLa cells at 48 h after transfection with the 9rR-LTVSPWY/pGL3 complex (N/P 6:1, black bars) or a vehicle control (blank, white bars). The data are shown as the means ± SD. All measurements were performed in triplicate. The “***” symbol indicates a significant difference between the two groups based on three independent experiments (p<0.001). The “#” symbol indicates no significant difference between the two groups based on three independent experiments (p>0.05).
Mentions: Because the confocal imaging and FACS results suggested that N/P = 6 results in the highest transfection efficiency, we conducted further experiments with the peptide/pDNA complexes prepared at an N/P ratio of 6. To quantify the specificity and efficiency of transfection in various types of cells, we performed quantitative analysis using flow cytometry. Figure 4A showed that the percentages of 5–8F, SKOV-3, MCF-7 and HeLa cells expressing GFP were approximately 30.33, 8.23, 1.04 and 1.05, respectively, implying that the plasmids were expressed in 5–8F and SKOV-3 cells, with significantly different expression levels in the experimental groups and the blank group, but not to MCF-7 or HeLa, which exhibited no significant difference between the experimental groups and the blank group. Next, we performed luciferase expression assays using the four cell lines. Figure 4B indicates that 9rR-LTVSPWY delivered the plasmid into 5–8F and SKOV-3 cells but not MCF-7 or HeLa cells, verifying the excellent selectivity of this method.

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