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. 2017 Oct 25;4(10):170822.
doi: 10.1098/rsos.170822. eCollection 2017 Oct.

Gene delivery ability of polyethylenimine and polyethylene glycol dual-functionalized nanographene oxide in 11 different cell lines

Liping Wu  1 , Jinshan Xie  1   2 , Tan Li  1 , Zihao Mai  1 , Lu Wang  1 , Xiaoping Wang  2 , Tongsheng Chen  1
Affiliations

Gene delivery ability of polyethylenimine and polyethylene glycol dual-functionalized nanographene oxide in 11 different cell lines

Liping Wu et al. R Soc Open Sci. .

Abstract

We recently developed a polyethylenimine (PEI) and polyethylene glycol (PEG) dual-functionalized reduced graphene oxide (GO) (PEG-nrGO-PEI, RGPP) for high-efficient gene delivery in HepG2 and Hela cell lines. To evaluate the feasibility and applicability of RGPP as a gene delivery carrier, we here assessed the transfection efficiency of RGPP on gene plasmids and siRNA in 11 different cell lines. Commercial polyalkyleneimine cation transfection reagent (TR) was used as comparison. In HepG2 cells, RGPP exhibited much stronger delivery ability for siRNA and large size plasmids than TR. For green fluorescent protein (GFP) plasmid, RGPP showed about 47.1% of transfection efficiency in primary rabbit articular chondrocytes, and about 27% of transfection efficiency in both SH-SY5Y and A549 cell lines. RGPP exhibited about 37.2% of GFP plasmid transfection efficiency in EMT6 cells and about 26.0% of GFP plasmid transfection efficiency in LO2 cells, but induced about 33% of cytotoxicity in both cell lines. In 4T1 and H9C2 cell lines, RGPP had less than 10% of GFP plasmid transfection efficiency. Collectively, RGPP is a potential nano-carrier for high-efficiency gene delivery, and needs to be further optimized for different cell lines.

Keywords: gene delivery; graphene oxide; polyethylene glycol; polyethylenimine; transfection efficiency.

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Conflict of interest statement

We declare we have no competing interests.

Figures

Figure 1.
Figure 1.
Transfection efficiency of RGPP on plasmids and siRNA in HepG2 cells. (a,b) Relative cell viability of cells treated with different concentrations of RGPP or TR for 48 h determined by CCK-8 assay. (c) Transfection efficiency of RGPP or TR for various plasmids after transfection for 48 h determined by FCM analysis. *p < 0.05 and **p < 0.01, compared with RGPP group. (d) Fluorescence microscopic images of cells exclusively transfected with GFP-Bak, GFP-Puma, GFP-Bax 1-2/L-6, GFP-Bcl-xL and GFP-NPM NLS1/2D plasmids by using RGPP at N/P ratio of 60 or 0.4% of TR for 48 h. Scale bar, 200 µm. (e) Transfection efficiency of RGPP or TR on siRNA after transfection for 4 h determined by FCM analysis. ***p < 0.001, compared with RGPP at N/P ratio of 5; ##p < 0.01, compared with 0.1% of TR. (f) Fluorescence microscopic images of cells transfected with siRNA by using RGPP at N/P ratio of 15 or 0.2% of TR for 4 h. Scale bar, 200 µm.
Figure 2.
Figure 2.
Transfection efficiency of RGPP on GFP plasmid in SH-SY5Y cells. (a,b) Relative cell viability of cells treated with different concentrations of RGPP or TR for 48 h determined by CCK-8 assay. (c) Transfection efficiency of RGPP or TR on GFP plasmid after transfection for 48 h determined by FCM analysis. **p < 0.01 and ***p < 0.001, compared with RGPP at N/P ratio of 15; #p < 0.05 and ##p < 0.01, compared with 0.2% of TR. (d) Fluorescence microscopic images of cells transfected with GFP plasmid by using RGPP at N/P ratio of 90 or 0.4% of TR for 48 h. Scale bar, 200 µm.
Figure 3.
Figure 3.
Transfection efficiency of RGPP on GFP plasmid in mouse cancer cell lines. (a,b) Relative cell viability of EMT6 cells treated with different concentrations of RGPP or TR for 48 h determined by CCK-8 assay. (c) Transfection efficiency of RGPP or TR on GFP plasmid after transfection for 48 h in EMT6 cells determined by FCM analysis. *p < 0.05 and ***p < 0.001, compared with RGPP at N/P ratio of 15; ##p < 0.01 and ###p < 0.001, compared with 0.2% of TR. (d) Fluorescence microscopic images of EMT6 cells transfected with GFP plasmid by using RGPP at N/P ratio of 60 or 0.6% of TR for 48 h. Scale bar, 200 µm. (e,f) Relative cell viability of 4T1 cells treated with different concentrations of RGPP or TR for 48 h determined by CCK-8 assay. (g) Transfection efficiency of RGPP or TR on GFP plasmid after transfection for 48 h in 4T1 cells determined by FCM analysis. (h) Fluorescence microscopic images of 4T1 cells transfected with GFP plasmid by using RGPP at N/P ratio of 30 or 0.6% of TR for 48 h. Scale bar, 200 µm.
Figure 4.
Figure 4.
Transfection efficiency of RGPP on GFP plasmid in LO2 cells and H9C2 cells. (a,b) Relative cell viability of LO2 cells treated with different concentrations of RGPP or TR for 48 h determined by CCK-8 assay. (c) Transfection efficiency of RGPP or TR on GFP plasmid after transfection for 48 h in LO2 cells determined by FCM analysis. *p < 0.05 and ***p < 0.001, compared with RGPP at N/P ratio of 15; #p < 0.05 and ##p < 0.01, compared with 0.2% of TR. (d,e) Relative cell viability of H9C2 cells treated with different concentrations of RGPP or TR for 48 h determined by CCK-8 assay. (f) Transfection efficiency of RGPP or TR on GFP plasmid after transfection for 48 h in H9C2 cells determined by FCM analysis. #p < 0.05 and ##p < 0.01, compared with 0.2% of TR.
Figure 5.
Figure 5.
Transfection efficiency of RGPP on GFP plasmid in primary rabbit articular chondrocyte. (a,b) Relative cell viability of rabbit articular chondrocyte treated with different concentrations of RGPP or TR for 48 h determined by CCK-8 assay. (c) Transfection efficiency of RGPP or TR on GFP plasmid after transfection for 48 h in rabbit articular chondrocyte determined by FCM analysis. **p < 0.01 and ***p < 0.001, compared with RGPP at N/P ratio of 15; ###p < 0.001, compared with 0.2% of TR.
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