Characterization of synergistic anti-tumor effects of doxorubicin and p53 via graphene oxide-polyethyleneimine nanocarriers

doi:10.1016/j.jmst.2017.05.005

Abstract

Abstract:

Co-delivery of chemical drugs and therapeutic genes for synergistic therapy provides a promising strategy to treat devastating diseases. However, the real-time coordination patterns between chemical drugs and therapeutic genes remain poorly understood. Herein, the complexes of doxorubicin/graphene oxide-polyethyleneimine/p53 plasmid (Dox/GO-PEI/p53) were fabricated and employed to investigate the synergistic manner between Dox and p53 in the inhibition of HeLa cell growth. GO was conjugated with PEI to form the GO-PEI backbone as the delivery vector. The GO backbone provided surfaces with a high specific area to load Dox via the π-π stacking interaction, and was able to release Dox significantly faster at pH 5.0 than at pH 7.0, while the positively charged PEI section of GO-PEI could condense plasmids into GO-PEI/DNA nanoparticles via the electrostatic interaction. The nanoparticles efficiently mediated the transfection of DNA in HeLa cells, with lower cytotoxicity compared to PEI/DNA nanoparticles. Furthermore, the complexes of Dox/GO-PEI/p53 released Dox and expressed p53 gene in a sequential manner, and showed successive inhibition of the in vitro growth of HeLa cells. This type of drug/GO-PEI/DNA complex can be employed as a platform to investigate the coordination pattern between chemical drugs and therapeutic genes for tumor therapy.

Key words: Graphene oxide, Polyethyleneimine, Doxorubicin, p53, Successive inhibition

Xie Bei, Yi Jipeng, Peng Jian, Zhang Xing, Lei Lei, Zhao Dapeng, Lei Zhixin, Nie Hemin. Characterization of synergistic anti-tumor effects of doxorubicin and p53 via graphene oxide-polyethyleneimine nanocarriers[J]. J. Mater. Sci. Technol., 2017, 33(8): 807-814.

Figures/Tables 8

Fig. 1. Characterization of different GO-PEI based complexes: (a) UV-vis spectroscopy of GO, PEI, GO-PEI, Dox, and Dox/GO-PEI; (b) FTIR analysis of GO and GO-PEI; (c) emission of Dox, GO and Dox/GO-PEI at 600 nm.

Fig. 2. Comparison of particle sizes (a) and zeta potentials (b) of GO, PEI, GO-PEI, Dox/GO-PEI, GO-PEI/p53 and Dox/GO-PEI/p53; TEM micrographs of GO (c), GO-PEI (d) and Dox/GO-PEI/p53 (e) (*p < 0.05).

Fig. 3. Photos showing colloidal stability of GO, GO/PEI mixture and GO-PEI conjugates in water, PBS, and 10% serum-containing mediumat 1 h and 3 h, respectively.

Fig. 4. Endocytosis of Dox/GO-PEI (a), and in vitro release profiles of Dox in the buffers with different pH values (5, 7, and 9) (b) (*p < 0.05).

Fig. 5. Simultaneous delivery of Dox and DNA: (a) agarose gel electrophoretic assay of GO-PEI/p53 complexes fabricated at different PEI/DNA ratios, 8:1, 16:1, 25:1, 40:1 and 70:1, with free native plasmid as control. Lane 1: naked p53; lanes 2-6: GO-PEI/p53 complexes fabricated at PEI/DNA ratios of 8:1, 16:1, 25:1, 40:1 and 70:1, respectively; (b) co-localization of green fluorescence of EGFP and red fluorescence of Dox delivered by Dox/GO-PEI/EGFP. Scale bar = 100 μm.

Fig. 6. Comparison of cell viability induced by GO, PEI, GO-PEI, Dox/GO-PEI, GO-PEI/p53, Dox/GO-PEI/p53 and free Dox (*p < 0.05).

Fig. 7. Real-time monitoring of the proliferation of HeLa cells treated by GO, PEI, GO-PEI, Dox/GO-PEI, GO-PEI/p53, Dox/GO-PEI/p53 and free Dox, with cell cultures without treatment as control.

Scheme 1 Schematic of co-delivery and sequential action of Dox and p53 gene in inhibition of HeLa cell growth via Dox/GO-PEI/p53 complexes.

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