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. 2019 Aug 9:14:6371-6385.
doi: 10.2147/IJN.S211744. eCollection 2019.

Inhibition of proliferation and migration of tumor cells through phenylboronic acid-functionalized polyamidoamine-mediated delivery of a therapeutic DNAzyme Dz13

Jiebing Yang #  1 , Jiayuan Zhang #  1 , Jiakai Xing  1 , Zhiyuan Shi  1 , Haobo Han  1 , Quanshun Li  1
Affiliations

Inhibition of proliferation and migration of tumor cells through phenylboronic acid-functionalized polyamidoamine-mediated delivery of a therapeutic DNAzyme Dz13

Jiebing Yang et al. Int J Nanomedicine. .

Erratum in

Abstract

Background: The phenylboronic acid-functionalized polyamidoamine (PP) was employed as a gene carrier for Dz13 delivery, inducing an obvious anticancer response.

Materials and methods: The Dz13 condensation ability of PP was evaluated through gel retardation assay. The cellular uptake mechanism of PP/Dz13 nanoparticles was studied using confocal laser scanning microscope and flow cytometer. The inhibition ability of cell proliferation, migration and invasion was investigated through MTT assay, flow cytometry, wound healing and Transwell migration assays, using hepatocarcinoma cell line HepG2 as a model. Finally, Western blotting analysis was used to detect the signaling pathway associated with the inhibition of cell apoptosis and migration induced by Dz13 delivery.

Results: The carrier PP could efficiently condense Dz13 into stable nanoparticles at mass ratios of >1.5. The hydrodynamic diameter and zeta potential of PP/Dz13 nanoparticles were measured to be 204.77 nm and +22.00 mV at a mass ratio of 10.0, respectively. The nanoparticles could realize an efficient cellular uptake in sialic acid-dependent endocytosis manner. Moreover, the nanoparticles exhibited an obvious antiproliferation effect through the induction of cell apoptosis and cell cycle arrest due to the cleavage of c-Jun mRNA. Besides, the suppression of cell migration and invasion could be achieved after the PP/Dz13 transfection, attributing to the decreased expression level of MMP-2 and MMP-9.

Conclusion: The PP provided a potential delivery system to achieve the tumor-targeting gene therapy.

Keywords: DNAzyme; antimigration effect; antiproliferation effect; gene therapy; phenylboronic acid; polyamidoamine.

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Conflict of interest statement

The authors report no conflicts of interest in this work.

Figures

Scheme 1
Scheme 1
The construction of phenylboronic acid-functionalized polyamidoamine (PP) and its application in Dz13 delivery. Abbreviation: PAMAM, polyamidoamine.
Figure 1
Figure 1
(A) The synthetic illustration of the gene carrier PP and (B) gel retardation assay for the Dz13 binding and condensation ability of PP and PAMAM. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine; EDC/NHS, 1-[3-(dimethylamino)propyl]-3-ethyl carbodiimine/N-hydroxysuccinimide; RT, room temperature.
Figure 2
Figure 2
The 1H NMR spectra of (A) PAMAM and (B) PP in D2O. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 3
Figure 3
The TEM images of PAMAM/Dz13 (A) and PP/Dz13 nanoparticles (B). The scale bar is 200 nm. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 4
Figure 4
The CLSM images of HepG2 cells transfected with PP/Dz13 and PAMAM/Dz13 nanoparticles with the pretreatment of 1 mM PBA for 1 hr. The scale bar is 20 μm. Abbreviations: CLSM, confocal laser scanning microscopy; PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 5
Figure 5
Endosomal escape of PP/Dz13 nanoparticles through CLSM. The scale bar is 10 μm. Abbreviations: CLSM, confocal laser scanning microscopy; PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 6
Figure 6
The inhibition of cell proliferation in HepG2 cells after the transfection of PP/Dz13 (mass ratio of 10) and PAMAM/Dz13 nanoparticles (mass ratio of 7.5). The concentrations of free PP and PAMAM were 40 and 30 μg/mL, respectively. The data were presented as mean value ± SD of triplicate measurements (n.s., not significant; *p<0.05 and ***p<0.001). Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 7
Figure 7
The cell apoptosis analysis of HepG2 cells after the transfection of PP/Dz13 and PAMAM/Dz13 nanoparticles: (A) control, (B) Dz13, (C) PP, (D) PP/Dz13Scr, (E) PAMAM/Dz13, (F) PAMAM, (G) PAMAM/Dz13Scr and (H) PP/Dz13 nanoparticles. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 8
Figure 8
Western blotting assay for the expression level of associated proteins in HepG2 cells after Dz13 transfection. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 9
Figure 9
Cell cycle arrest (A) and the relative cell cycle phase (B) in HepG2 cells after Dz13 transfection: (a) control, (b) Dz13, (c) PP, (d) PP/Dz13Scr, (e) PP/Dz13 and (f) PAMAM/Dz13 nanoparticles. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 10
Figure 10
The antimigration assay in HepG2 cells after Dz13 transfection for different incubation times (A) and the quantitative wound size (B). The data were presented as mean value ± SD of triplicate experiments (*p<0.05). The scale bar is 200 μm. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
Figure 11
Figure 11
Transwell migration assay of HepG2 cells after the carriers-mediated Dz13 transfection: (A) control, (B) Dz13, (C) PP, (D) PP/Dz13Scr, (E) PP/Dz13 and (F) PAMAM/Dz13 nanoparticles. The scale bar is 50 μm. Abbreviations: PP, phenylboronic acid-functionalized polyamidoamine; PAMAM, polyamidoamine.
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