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

Varying efficiency of SW-induced siRNA delivery in different cell lines.(a and b) Human smooth muscle cells (HSMCs) or murine colon adenocarcinoma (CT26) cells were transfected with Cy3-labeled GAPDH siRNAs by SW treatment (0.04 mJ/mm2) or Lipofectamine (positive control). Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled GAPDH siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus the control group without SW treatment. Error bars represent SD.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Varying efficiency of SW-induced siRNA delivery in different cell lines.(a and b) Human smooth muscle cells (HSMCs) or murine colon adenocarcinoma (CT26) cells were transfected with Cy3-labeled GAPDH siRNAs by SW treatment (0.04 mJ/mm2) or Lipofectamine (positive control). Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled GAPDH siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus the control group without SW treatment. Error bars represent SD.

Mentions: At 24 h post-SW treatment, HSMCs and CT26 cells showed similar siRNA transfection efficiency to Lipofectamine treatment (Fig. 2a,b). The mean GAPDH protein expression was significantly lower in cells transfected with GAPDH siRNA by SW than in the control groups (p < 0.05). Additional cell lines, including a human prostate cancer cell line (PC-3), immortalized mouse aortic endothelial cells (iMAEC) and monkey kidney fibroblast (COS-7) cells were transfected with Cy3-labeled VEGF, KDR and GAPDH siRNAs by SW treatment (Supplementary Fig. 1). The data suggest that siRNAs were delivered into cells by low-energy SW treatment with suppression of the target gene expression.


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)

Varying efficiency of SW-induced siRNA delivery in different cell lines.(a and b) Human smooth muscle cells (HSMCs) or murine colon adenocarcinoma (CT26) cells were transfected with Cy3-labeled GAPDH siRNAs by SW treatment (0.04 mJ/mm2) or Lipofectamine (positive control). Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled GAPDH siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus the control group without SW treatment. Error bars represent SD.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Varying efficiency of SW-induced siRNA delivery in different cell lines.(a and b) Human smooth muscle cells (HSMCs) or murine colon adenocarcinoma (CT26) cells were transfected with Cy3-labeled GAPDH siRNAs by SW treatment (0.04 mJ/mm2) or Lipofectamine (positive control). Cells were fixed, and transfection of siRNAs or the vector was visualized by fluorescence microscopy. Cy3-labeled GAPDH siRNA immunofluorescence staining is indicated in red and DAPI-stained nuclei in blue. Representative immunoblots and quantitative data are shown (n = 3). *p < 0.05 versus the control group without SW treatment. Error bars represent SD.
Mentions: At 24 h post-SW treatment, HSMCs and CT26 cells showed similar siRNA transfection efficiency to Lipofectamine treatment (Fig. 2a,b). The mean GAPDH protein expression was significantly lower in cells transfected with GAPDH siRNA by SW than in the control groups (p < 0.05). Additional cell lines, including a human prostate cancer cell line (PC-3), immortalized mouse aortic endothelial cells (iMAEC) and monkey kidney fibroblast (COS-7) cells were transfected with Cy3-labeled VEGF, KDR and GAPDH siRNAs by SW treatment (Supplementary Fig. 1). The data suggest that siRNAs were delivered into cells by low-energy SW treatment with suppression of the target gene expression.

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