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. 2020 Jul 20;15(1):150.
doi: 10.1186/s11671-020-03377-y.

Gint4.T-Modified DNA Tetrahedrons Loaded with Doxorubicin Inhibits Glioma Cell Proliferation by Targeting PDGFRβ

Feng Wang  1 , Yanghao Zhou  2 , Si Cheng  3 , Jinhe Lou  4 , Xiang Zhang  2 , Qiuguang He  2 , Ning Huang  2 , Yuan Cheng  5
Affiliations

Gint4.T-Modified DNA Tetrahedrons Loaded with Doxorubicin Inhibits Glioma Cell Proliferation by Targeting PDGFRβ

Feng Wang et al. Nanoscale Res Lett. .

Abstract

Glioma is one of the deadliest intrinsic brain tumours due to its invasive growth. The effect of glioma treatment is poor because of the presence of the blood-brain barrier and blood tumour barrier and insufficient drug targeting. DNA tetrahedrons (TDN) show great potential for drug delivery and may be a novel therapeutic strategy for glioma. In this study, we used TDN to deliver doxorubicin (DOX) for the glioma therapy. Gint4.T, an aptamer that could recognize platelet-derived growth factor receptor β on tumour cell, was used to modify TDN (Apt-TDN) for targeted drug delivery. The TDN were self-assembled by one-step synthesis, which showed small size (10 nm) and negative charge. Fetal bovine serum test showed its stability as a drug delivery vehicle. Apt-TDN could be effectively taken up by U87MG cells. Compared with DOX and DOX@TDN (TDN loaded with DOX), the DOX@Apt-TDN (Gint4.T-modified TDN loaded with DOX) showed more early apoptosis rate, higher cell cycle arrest, and greater cytotoxicity towards U87MG cells. In conclusion, our findings indicated that DOX@Apt-TDN provides a novel therapy with promising clinical application for gliomas patients.

Keywords: DNA tetrahedron; Gint4.T; Glioma; Nanostructures; Platelet-derived growth factor receptor β.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Fig. 1
Fig. 1
a Synthesis of the DNA tetrahedron and Gint4.T-TDN. Lane 1: S1; lane 2: S1+S2; lane 3: S1+S2+S3; lane 4: S1+S2+S3+S4 (TDN); lane 5: TDN mixed with Apt-tail (Gint4.T); Apt-TDN. Lane 1 was not visible because nucleic acid dyes cannot properly stain single-strand DNA. b AFM images showed that the heights of the TDN and Apt-TDN were ~ 2 nm. c Determination of the particle size and zeta potential of the TDN and Apt-TDN by dynamic light scattering (DLS). The average particle sizes of the TDN and Apt-TDN were 10.10 nm (A) and 13.54 nm (B), respectively. The average zeta potentials of the TDN and Apt-TDN were − 5.69 mV (C) and − 7.3 mV (D), respectively
Fig. 2
Fig. 2
a Gel electrophoresis showed that the TDN remained stable for 24 h in complete medium at 37 °C (A); the TDN remained stable for 7 h when the concentration of foetal bovine serum was increased to 50% (B). b U87MG cells were co-cultured with the TDN at different concentrations (10–500 nM) for 24 h and 48 h. The CCK-8 assay showed that the activity of the U87MG cells was not affected, which indicated the biosafety of the TDN
Fig. 3
Fig. 3
a A standard curve of DOX concentrations in PBS buffer; λex = 480 nm and λem = 590 nm. The amount of DOX carried by the TDN and Apt-TDN. b DOX intercalated into the double-strand DNA of the TDN and Apt-TDN. When the DOX concentration reached 14 μM and the intercalated DOX concentration in the TDN and Apt-TDN peaked at 5.5 μM and 6.0 μM, respectively, a single DNA tetrahedron could carry 55 Dox molecules, while a single aptamer-modified DNA tetrahedron carried 60 DOX molecules. c Fluorescence spectra of DOX in the supernatant. Doxorubicin was mixed with the TDN at increasing molar ratios (0, 0.0005, 0.001, 0.005, 0.01, and 0.05 from top to bottom). When the molar ratio was 1:20, the fluorescence was quenched
Fig. 4
Fig. 4
a U87MG cell uptake of the TDN and Apt-TDN (TDN-Gint4.T). The TDN entered U87MG cells directly without transfection agents, and the uptake of Apt-TDN (linked to the aptamer Gint4.T) was significantly increased and competitively inhibited by free Apt (Gint4.T), indicating that the aptamer Gint4.T plays a significant role in cellular targeting. The scale bar represents 50 μm. b Flow cytometry curves show the intracellular uptake of the TDN, Apt-TDN, and Apt-TDN+Apt after incubation for 3 h
Fig. 5
Fig. 5
a Cellular uptake of DOX, the DOX@TDN, and the DOX@Apt-TDN. Modified with the aptamer Gint4.T, the Apt-TDN could deliver more doxorubicin to U87MG cells than the TDN. In addition, the TDN could carry more drug to cells than the drug alone. The scale bars indicate 50 μm. b Semi-quantitative analysis of the fluorescence intensity of doxorubicin with PBS, DOX, DOX@TDN, and DOX@Apt-TDN treatment (compared to blank: *p < 0.05, **p < 0.01)
Fig. 6
Fig. 6
a Cytotoxicity of DOX, the DOX@TDN, and the DOX@Apt-TDN at various concentrations. The inhibition rate of the U87MG cells was significantly increased with increasing DOX concentration, but the DOX@TDN and DOX@Apt-TDN groups exhibited significantly increased cytotoxicity compared with the DOX group. The cell inhibition rate of the DOX@Apt-TDN group was also significantly higher than that of the DOX@TDN group (compared to DOX, **p < 0.05; compared to DOX, ***p < 0.01; compared to DOX@Apt-TDN, #p < 0.05). b Apoptosis of U87MG cells after incubation with PBS, DOX, the DOX@TDN, and the DOX@Apt-TDN for 24 h. c Flow cytometry histograms of the U87MG cell cycle after incubation with PBS, DOX, the DOX@TDN, and the DOX@Apt-TDN for 24 h (compared to control, **p < 0.05; compared to control, ***p < 0.01; compared to DOX@Apt-TDN, #p < 0.01)
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