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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 of VEGF siRNAs ex vivo and in vivo.(a and b) Mouse aorta rings were isolated. VEGFR2 siRNAs were added to the culture medium of aortic rings and treated with SW at 0.045 mJ/mm2, incubated for 6bdays, at which point an aorta ring assay for ex vivo angiogenesis was performed. VEGF increased the number of microvessels sprouting from the aortic rings. Representative images are shown (n = 4). *p < 0.05 versus the control group (no SW treatment). Error bars represent SD. (c) Cy3-labeled VEGF siRNA following SW treatment (0.02 mJ/mm2) was transfected into CT26 tumors. (d and e) Data represent the expression levels of VEGF in CT26 tumors following SW treatment and VEGF siRNA. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus + VEGF without SW treatment. Error bars represent SD. (f and g) SW treatment (0.02 mJ/mm2) resulted in weak CD31 staining.
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f4: SW-induced transfection of VEGF siRNAs ex vivo and in vivo.(a and b) Mouse aorta rings were isolated. VEGFR2 siRNAs were added to the culture medium of aortic rings and treated with SW at 0.045 mJ/mm2, incubated for 6bdays, at which point an aorta ring assay for ex vivo angiogenesis was performed. VEGF increased the number of microvessels sprouting from the aortic rings. Representative images are shown (n = 4). *p < 0.05 versus the control group (no SW treatment). Error bars represent SD. (c) Cy3-labeled VEGF siRNA following SW treatment (0.02 mJ/mm2) was transfected into CT26 tumors. (d and e) Data represent the expression levels of VEGF in CT26 tumors following SW treatment and VEGF siRNA. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus + VEGF without SW treatment. Error bars represent SD. (f and g) SW treatment (0.02 mJ/mm2) resulted in weak CD31 staining.

Mentions: To define the efficiency of SW-induced siRNA transfection in intact vessels, an aorta ring assay was conducted for ex vivo angiogenesis3233. VEGF increased the number of microvessels sprouting from aortic rings isolated from mice, and transfection of VEGF siRNA by SW treatment markedly inhibited the sprouting of VGEF-induced microvessels (Fig. 4a,b). These data suggest that siRNAs was successfully introduced into the cells and functionally worked well in ex vivo by SW-induced transfection


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 of VEGF siRNAs ex vivo and in vivo.(a and b) Mouse aorta rings were isolated. VEGFR2 siRNAs were added to the culture medium of aortic rings and treated with SW at 0.045 mJ/mm2, incubated for 6bdays, at which point an aorta ring assay for ex vivo angiogenesis was performed. VEGF increased the number of microvessels sprouting from the aortic rings. Representative images are shown (n = 4). *p < 0.05 versus the control group (no SW treatment). Error bars represent SD. (c) Cy3-labeled VEGF siRNA following SW treatment (0.02 mJ/mm2) was transfected into CT26 tumors. (d and e) Data represent the expression levels of VEGF in CT26 tumors following SW treatment and VEGF siRNA. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus + VEGF without SW treatment. Error bars represent SD. (f and g) SW treatment (0.02 mJ/mm2) resulted in weak CD31 staining.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: SW-induced transfection of VEGF siRNAs ex vivo and in vivo.(a and b) Mouse aorta rings were isolated. VEGFR2 siRNAs were added to the culture medium of aortic rings and treated with SW at 0.045 mJ/mm2, incubated for 6bdays, at which point an aorta ring assay for ex vivo angiogenesis was performed. VEGF increased the number of microvessels sprouting from the aortic rings. Representative images are shown (n = 4). *p < 0.05 versus the control group (no SW treatment). Error bars represent SD. (c) Cy3-labeled VEGF siRNA following SW treatment (0.02 mJ/mm2) was transfected into CT26 tumors. (d and e) Data represent the expression levels of VEGF in CT26 tumors following SW treatment and VEGF siRNA. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus + VEGF without SW treatment. Error bars represent SD. (f and g) SW treatment (0.02 mJ/mm2) resulted in weak CD31 staining.
Mentions: To define the efficiency of SW-induced siRNA transfection in intact vessels, an aorta ring assay was conducted for ex vivo angiogenesis3233. VEGF increased the number of microvessels sprouting from aortic rings isolated from mice, and transfection of VEGF siRNA by SW treatment markedly inhibited the sprouting of VGEF-induced microvessels (Fig. 4a,b). These data suggest that siRNAs was successfully introduced into the cells and functionally worked well in ex vivo by SW-induced transfection

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