Limits...
Novel mechanism of gene transfection by low-energy shock wave.

Ha CH, Lee SC, Kim S, Chung J, Bae H, Kwon K - Sci Rep (2015)

Bottom Line: Furthermore SW-induced siRNA transfection was not mediated by SW-induced sonoporation, but by microparticles (MPs) secreted from the cells.Interestingly, the transfection effect of the siRNAs was transferable through the secreted MPs from human umbilical vein endothelial cell (HUVEC) culture medium after treatment with SW, into HUVECs in another culture plate without SW treatment.In this study, we suggest for the first time a mechanism of gene transfection induced by low-energy SW through secreted MPs, and show that it is an efficient physical gene transfection method in vitro and represents a safe therapeutic strategy for site-specific gene delivery in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Asan Institute for Life Sciences, Asan Medical Center, College of Medicine, University of Ulsan, 86 Asanbyeoungwon-gil, Songpa-gu, Seoul, 138-736, Korea.

ABSTRACT
Extracorporeal shock wave (SW) therapy has been studied in the transfection of naked nucleic acids into various cell lines through the process of sonoporation, a process that affects the permeation of cell membranes, which can be an effect of cavitation. In this study, siRNAs were efficiently transfected into primary cultured cells and mouse tumor tissue via SW treatment. Furthermore SW-induced siRNA transfection was not mediated by SW-induced sonoporation, but by microparticles (MPs) secreted from the cells. Interestingly, the transfection effect of the siRNAs was transferable through the secreted MPs from human umbilical vein endothelial cell (HUVEC) culture medium after treatment with SW, into HUVECs in another culture plate without SW treatment. In this study, we suggest for the first time a mechanism of gene transfection induced by low-energy SW through secreted MPs, and show that it is an efficient physical gene transfection method in vitro and represents a safe therapeutic strategy for site-specific gene delivery in vivo.

No MeSH data available.


Related in: MedlinePlus

SW-induced transfection without sonoporation.(a) Cy3-labeled VEGFR2 siRNAs were added to HUVEC culture medium and treated with SW at 0.045 mJ/mm2 prior to incubation for 0 min–48 h. (b) SW-stimulated HUVECs were incubated for the indicated times followed by a medium change to remove siRNAs and were then incubated for an additional 24 h. (c) Cy3-labeled VEGFR2 siRNAs were added to confluent HUVECs in the absence or presence of SW and incubated for 24 h. After incubation, control and SW-treated HUVEC media were transferred to HUVECs without SW treatment followed by an additional 24-h incubation. Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled VEGFR-2 siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4525295&req=5

f3: SW-induced transfection without sonoporation.(a) Cy3-labeled VEGFR2 siRNAs were added to HUVEC culture medium and treated with SW at 0.045 mJ/mm2 prior to incubation for 0 min–48 h. (b) SW-stimulated HUVECs were incubated for the indicated times followed by a medium change to remove siRNAs and were then incubated for an additional 24 h. (c) Cy3-labeled VEGFR2 siRNAs were added to confluent HUVECs in the absence or presence of SW and incubated for 24 h. After incubation, control and SW-treated HUVEC media were transferred to HUVECs without SW treatment followed by an additional 24-h incubation. Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled VEGFR-2 siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue.

Mentions: To confirm whether SW-induced siRNA transfection was caused by sonoporation, Cy3-labeled VEGFR2 siRNAs were added to HUVEC culture medium for transfection and treated with SW (0.04 mJ/mm2), followed by incubation for 0 min to 48 h (Fig. 3).


Novel mechanism of gene transfection by low-energy shock wave.

Ha CH, Lee SC, Kim S, Chung J, Bae H, Kwon K - Sci Rep (2015)

SW-induced transfection without sonoporation.(a) Cy3-labeled VEGFR2 siRNAs were added to HUVEC culture medium and treated with SW at 0.045 mJ/mm2 prior to incubation for 0 min–48 h. (b) SW-stimulated HUVECs were incubated for the indicated times followed by a medium change to remove siRNAs and were then incubated for an additional 24 h. (c) Cy3-labeled VEGFR2 siRNAs were added to confluent HUVECs in the absence or presence of SW and incubated for 24 h. After incubation, control and SW-treated HUVEC media were transferred to HUVECs without SW treatment followed by an additional 24-h incubation. Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled VEGFR-2 siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525295&req=5

f3: SW-induced transfection without sonoporation.(a) Cy3-labeled VEGFR2 siRNAs were added to HUVEC culture medium and treated with SW at 0.045 mJ/mm2 prior to incubation for 0 min–48 h. (b) SW-stimulated HUVECs were incubated for the indicated times followed by a medium change to remove siRNAs and were then incubated for an additional 24 h. (c) Cy3-labeled VEGFR2 siRNAs were added to confluent HUVECs in the absence or presence of SW and incubated for 24 h. After incubation, control and SW-treated HUVEC media were transferred to HUVECs without SW treatment followed by an additional 24-h incubation. Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled VEGFR-2 siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue.
Mentions: To confirm whether SW-induced siRNA transfection was caused by sonoporation, Cy3-labeled VEGFR2 siRNAs were added to HUVEC culture medium for transfection and treated with SW (0.04 mJ/mm2), followed by incubation for 0 min to 48 h (Fig. 3).

Bottom Line: Furthermore SW-induced siRNA transfection was not mediated by SW-induced sonoporation, but by microparticles (MPs) secreted from the cells.Interestingly, the transfection effect of the siRNAs was transferable through the secreted MPs from human umbilical vein endothelial cell (HUVEC) culture medium after treatment with SW, into HUVECs in another culture plate without SW treatment.In this study, we suggest for the first time a mechanism of gene transfection induced by low-energy SW through secreted MPs, and show that it is an efficient physical gene transfection method in vitro and represents a safe therapeutic strategy for site-specific gene delivery in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Asan Institute for Life Sciences, Asan Medical Center, College of Medicine, University of Ulsan, 86 Asanbyeoungwon-gil, Songpa-gu, Seoul, 138-736, Korea.

ABSTRACT
Extracorporeal shock wave (SW) therapy has been studied in the transfection of naked nucleic acids into various cell lines through the process of sonoporation, a process that affects the permeation of cell membranes, which can be an effect of cavitation. In this study, siRNAs were efficiently transfected into primary cultured cells and mouse tumor tissue via SW treatment. Furthermore SW-induced siRNA transfection was not mediated by SW-induced sonoporation, but by microparticles (MPs) secreted from the cells. Interestingly, the transfection effect of the siRNAs was transferable through the secreted MPs from human umbilical vein endothelial cell (HUVEC) culture medium after treatment with SW, into HUVECs in another culture plate without SW treatment. In this study, we suggest for the first time a mechanism of gene transfection induced by low-energy SW through secreted MPs, and show that it is an efficient physical gene transfection method in vitro and represents a safe therapeutic strategy for site-specific gene delivery in vivo.

No MeSH data available.


Related in: MedlinePlus